Indolocarbazole derivatives that stimulate platelet production

ABSTRACT

Disclosed is an indolocarbazole derivative represented by the following formula (I), or a pharmaceutically acceptable salt thereof: ##STR1## [wherein R 1  represents substituted or unsubstituted lower alkyl, R 2  represents hydrogen, substituted or unsubstituted lower alkyl, R 3  and R 4  are the same or different, and represent hydrogen, nitro, NR 5  R 6 , OR 9 , substituted or unsubstituted lower alkyl, substituted or unsubstituted lower alkenyl, or CH═NNR 10  R 11 , W represents two hydrogen atoms or oxygen, and Me represents methyl, provided that when R 1  is unsubstituted lower alkyl, R 3  and R 4  are not simultaneously hydrogen.

BACKGROUND OF THE INVENTION

The present invention relates to a novel indolocarbazole derivative or apharmaceutically acceptable salt thereof. These compounds are expectedto be useful as a therapeutic agent for thrombocytopenia.

The decrease in the number of blood platelets due to various kinds ofhematopoietic disorders causes grave symptoms including an increasedtendency to hemorrhage. At present, platelet transfusion is effectiveagainst such decreases. However, a sufficient amount of blood plateletsis not always supplied. In addition to platelet transfusion, interleukin(IL) 6 and IL 11, which are hematopoietic factors that stimulate theproduction of blood platelets are known [Blood, 75, 1602 (1990); Blood,81, 901 (1993)].

Indolocarbazole derivatives having staurosporine skeleton are known tohave protein kinase C inhibitory activity, anti-tumor activity andantibacterial activity [WO89/07105, WO92/17181, Japanese PublishedUnexamined Patent Application No. 220196/87, Japanese PublishedUnexamined Patent Application No. 86068/93], platelet aggregationinhibitory activity [Japanese Published Unexamined Patent ApplicationNo. 72485/91, Japanese Published Unexamined Patent Application No.163325/91, Japanese Published Unexamined Patent Application No.220194/91, Japanese Published Unexamined Patent Application No.145085/92, Japanese Published Unexamined Patent Application No.364186/92], vasodilating activity [Japanese Published Unexamined PatentApplication No. 143877/89, Japanese Published Unexamined PatentApplication No. 9819/90], antiobestic activity [WO89/11295] ormacrophage activating activity [Journal of Antibiotics, 42, 1619 (1989)]and so on.

However, it is not known that indolocarbazole derivatives have anactivity to stimulate the production of blood platelets.

Most of the known indolocarbazole derivatives have hydrogen at theposition corresponding to R¹ in the following formula (I).Alternatively, the known indolocarbazole derivatives have lower alkyl atthe position corresponding to R¹, and hydrogen at the positionscorresponding to R³ and R⁴ in the following formula (I).

SUMMARY OF THE INVENTION

It is an object of the invention to provide a novel indolocarbazolederivative or a pharmaceutically acceptable salt thereof, which isuseful as a therapeutic agent for thrombocytopenia.

The present invention relates to an indolocarbazole derivativerepresented by the formula (I): ##STR2## [wherein R¹ representssubstituted or unsubstituted lower alkyl, R² represents hydrogen,substituted or unsubstituted lower alkyl, R³ and R⁴ are the same ordifferent, and represent hydrogen, nitro, NR⁵ R⁶ [wherein R⁵ and R⁶ arethe same or different and represent hydrogen, lower alkyl, an amino acidresidue where hydroxy group in the carboxylic acid is removed from theamino acid, alkanoyl or CO(CH₂)_(m) NR⁷ R⁸ (wherein m is an integer of 0to 3, R⁷ and R⁸ are the same or different and represent hydrogen orlower alkyl, or R⁷ and R⁸ are combined together with nitrogen atom toform a heterocyclic ring)], OR⁹ [wherein R⁹ represents long-chainedalkyl, alkanoyl, COCH₂ (OCH₂ CH₂)_(k) OCH₃ (wherein k is an integer of 1to 6), or CONR⁷ R⁸ (wherein R⁷ and R⁸ have the same meanings as definedabove)], substituted or unsubstituted lower alkyl, substituted orunsubstituted lower alkenyl, or CH═NNR¹⁰ R¹¹ (wherein R¹⁰ and R¹¹ arethe same or different and represent hydrogen, lower alkyl or aheterocyclic ring containing nitrogen atom), W¹ and W² are both hydrogenor are combined together to represent oxygen, and Me represents methyl,provided that when R¹ is unsubstituted lower alkyl, R³ and R⁴ are notsimultaneously hydrogen], or a pharmaceutically acceptable salt thereof.

The compound represented by the formula (I) is hereinafter referred toas to Compound (I). The same numbering applies to the compounds havingother formula numbers.

DETAILED DESCRIPTION OF THE INVENTION

The substituted or unsubstituted lower alkyl means a straight orbranched alkyl having 1-6 carbon atoms such as methyl, ethyl, propyl,isopropyl, butyl, isobutyl, secbutyl, tert-butyl, pentyl, neopentyl andhexyl, which is optionally substituted with 1 to 3 substituentsindependently selected from the group consisting of loweralkoxycarbonyl, OR¹² [wherein R¹² represents hydrogen, or lower alkylwhich may be substituted by NR^(7a) R^(8a) (wherein R^(7a) and R^(8a)have the same meanings as those defined for above-mentioned R⁷ and R⁸)],and SR¹³ [wherein, R¹³ represents hydrogen, heteroaryl group or loweralkyl which may be substituted by NR^(7b) R^(8b) (wherein, R^(7b) andR^(8b) have the same meanings as those defined for the above-mentionedR⁷ and R⁸)]. The alkyl moiety in the lower alkoxycarbonyl, and the alkylin the definitions of R¹² and R¹³ have the same definition as the loweralkyl mentioned above. The heteroaryl represents pyrrolyl, imidazolyl,thienyl, furyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl, pyridyl,pyrimidinyl, indolyl, quinolyl, and the like. The long-chained alkylmeans a linear or branched alkyl having 7-20 carbon atoms such asheptyl, octyl, isooctyl, nonyl, isononyl, pentadecanyl and eicosyl andthe like.

The substituted or unsubstituted lower alkenyl means an alkenyl having2-6 carbon atoms such as vinyl, allyl, butenyl pentenyl, hexenyl,pentadienyl, hexadienyl and the like, which is optionally substitutedwith 1 to 2 of the same substituents as those for the substituent in thelower alkyl mentioned above.

Alkanoyl means a straight or branched alkanoyl having 1-20 carbon atoms,such as formyl, acetyl, propionyl, isopropionyl, butyryl, valeryl,pivaloyl, hexanoyl, octanoyl, nonanoyl, palmitoyl, stearoyl and thelike.

Amino acid means glycine, alanine, serine, cysteine, lysine, glutamicacid, phenylalanine, proline and the like.

The heterocyclic group formed together with nitrogen atom and theheterocyclic group containing nitrogen atom are the same or differentand include pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl,pyrazolidinyl, pyrazolinyl, piperidyl, piperazinyl, morpholinyl,thiomorpholinyl, N-methylpiperazinyl, indolinyl, isoindolinyl and thelike.

In Compound (I), R¹ preferably represents unsubstituted alkyl, morepreferably methyl. R² preferably represents hydrogen. R³ and R⁴preferably represents NR⁵ R⁶ (wherein R⁵ and R⁶ have the same meaningsas defined above), OR⁹ (wherein R⁹ has the same meaning as definedabove), substituted or unsubstituted lower alkyl, substituted orunsubstituted lower alkenyl, or CH═NNR¹⁰ R¹¹ (wherein R¹⁰ and R¹¹ havethe same meanings as defined above). W preferably represents twohydrogen atoms.

The pharmaceutically acceptable salts of Compound (I) includepharmaceutically acceptable acid addition salts, which include saltswith inorganic acids such as hydrochloride, sulfate, and phosphate, andsalts with organic acids such as acetate, maleate, fumarate, tartrate,citrate, lactate, aspartate and glutamate.

The compounds of the present invention are normally obtained fromoptically active staurosporine as a starting material [Journal ofAntibiotics, 30, 275 (1977)]. The present invention covers all thepossible stereoisomers and a mixture thereof.

The processes for the production of Compound (I) are described below.

In the following production processes, when the defined group changesunder the condition of the process employed, or is inappropriate forcarrying out the process, typical protection and deprotection processesof the organic synthetic chemistry can be used to obtain the desiredcompound [see, e.g. T. W.Greene, Protective Groups in Organic Synthesis,John Wiley & Sons Inc. (1981)]. It is also possible to change the orderof the substituent introduction steps if necessary.

In the formulae and in the tables, Me, Et, t-Bu and Ac stand for methyl,ethyl, t-butyl and acetyl groups, respectively.

Production process 1

Compound (IA) [Compound (I) in which a substituent is introduced at R¹position] can be produced according to the following step. ##STR3## (Inthe formulae, R¹, W¹ and W² have the same meanings as defined above, andCbz represents benzyloxycarbonyl.)

Step 1

Compound (A) which is obtained by a known method (WO89-07105) fromstaurosporine [Journal of Antibiotics, 30, 275 (1977)] is allowed toreact with Compound (II) which is represented by the following formula(II):

    R.sup.1 Hal                                                (II)

(wherein, R¹ has the same definition as defined above, Hal representschloride, bromide or iodide), in the presence of a base in a solventwhich is inert to the reaction, and then the protecting group Cbz isremoved to give Compound (IA).

N,N-dimethylformamide (DMF), tetrahydrofuran (THF), toluene and the likecan be used alone or in combination as a reaction solvent, and sodiumhydroxide, potassium tert-butoxide and the like can be used as a base.The reaction is carried out at -20° to 50° C. and is completed in 1 to24 hours.

The protecting group Cbz is removed by catalytic reduction in a solventsuch as DMF and THF in the presence of a catalyst such as 10% Pd/C and20% Pd(OH)₂ /C in a hydrogen stream to give Compound (IA). The amount ofthe reducing catalyst to be used is 10-100% (by weight) based onCBz-protected derivative. The reaction is carried out at -10° to 50° C.and is completed in 1 to 24 hours.

Production process 2

Compound (IB) [Compound (I) in which a substituent is introduced at theR² position] can be produced according to the following steps, fromCompound (IA) which is obtained by the production process 1. ##STR4##(In the formulae, R^(2a) represents all the groups but hydrogen in thedefinition of R², and R¹, W¹ and W² have the same meanings as definedabove.)

Step 2

Compound (IA) is allowed to react with Compound (III) which isrepresented by the following formula (III):

    R.sup.14 CHO                                               (III)

(wherein, R¹⁴ represents substituted or unsubstituted alkyl having thenumber of carbon atoms which is less by one than that of the lower alkylin the definition of the above-defined R²), or a dimer thereof, in thepresence of a reducing agent such as sodium cyanoborohydride in asolvent such as a mixture of THF and water (10/1), at pH 5 to 6 to giveCompound (IB). Sodium cyanoborohydride, Compound (III) or the dimerthereof are used in amounts of 1 to 1.5 equivalents, respectively, basedon Compound (IA). The reaction is carried out at -10° to 50° C. and iscompleted in 1 to 8 hours.

Production process 3

Compound (IC) [Compound (I) in which a substituent is introduced at R³and/or R⁴ positions] can be produced according to the following steps oraccording to the process described in WO88/07045, from Compound (B)which is obtained by Production process 1 or Production process 2.##STR5## (Wherein R¹, R², W¹ and W² have the same meanings as definedabove, and the definitions of R^(3a) and R^(4a) are given in thefollowing steps.)

Step 3-1

(In the formulae, at least one of R^(3a) and R^(4a) is nitro.)

Compound (B) is allowed to react with fuming nitric acid in the presenceof trifluorosulfonic acid in a solvent such as dichloromethane to giveCompound (IC). Trifluorosulfonic acid and fuming nitric acid are used inamounts of 1 to 3 equivalents, respectively, based on Compound (B). Thereaction is carried out at -78° to 30° C. and is completed in 1 to 8hours.

Step 3-2

(In the formulae, at least one of R^(3a) and R^(4a) is hydroxymethyl.)

Compound (B) is allowed to react with dichloromethyl methyl ether in thepresence of a Lewis acid such as titanium tetrachloride in a solventsuch as methylene chloride, chloroform, and dichloroethane to giveformyl derivative which is a compound wherein at least one of R^(3a) andR^(4a) is formyl. Titanium tetrachloride and dichloromethyl methyl etherare used in amounts of 1 to 10 equivalents, respectively, based onCompound (B). The reaction is carried out at -10° to 80° C. and iscompleted in 1 to 8 hours.

The resulting formyl derivative is allowed to react with a reducingagent such as sodium borohydride in a solvent such as a mixed solvent ofTHF and methanol (1/1) to give Compound (1C). The sodium borohydride isused in an amount of 1 to 5 equivalents based on the formyl derivative.The reaction is carried out at -10° to 50° C. and is completed in 1 to 8hours.

Step 3-3

(In the formulae, at least one of R^(3a) and R^(4a) is substituted orunsubstituted lower alkenyl.)

The formyl derivative obtained as a reaction intermediate in theabove-mentioned step 3-2 is allowed to react with a correspondingWittig's reagent in a solvent such as dichloroethane to give Compound(IC). Wittig's reagent is used in an amount of 1 to 3 equivalents, basedon the formyl derivative. The reaction is carried out at 0° to 80° C.and is completed in 5 to 24 hours.

Step 3-4

(In the formulae, at least one of R^(3a) and R^(4a) is alkanoyloxy orCOCH₂ (OCH₂ CH₂)_(k) OCH₃ (wherein k has the same meaning as definedabove)).

The formyl derivative obtained as a reaction intermediate in theabove-mentioned step 3-2 is allowed to react with a peroxide such asm-chloroperbenzoic acid in the presence of a base such as sodiumhydrogen carbonate in a solvent such as dichloromethane and chloroformto give formic ester. The peroxide and the base are used in amounts of1-5 equivalents, and 3 to 10 equivalents, respectively, based on theformic ester. The reaction is carried out at -10° to 50° C. and iscompleted in 1 to 24 hours.

Then the obtained formic ester is allowed to react with a base such assodium methoxide, in a solvent such as dichloromethane to give hydroxyderivative. Sodium methoxide is used in an amount of 1 to 5 equivalents,based on the formic ester. The reaction is carried out at -10° to 50° C.and is completed in 10 minutes to 1 hour.

Then the above-mentioned hydroxy derivative is allowed to react with anacid halide such as chloride and bromide of a corresponding carboxylicacid in the presence of a base such as triethylamine in a solvent suchas dichloromethane to give Compound (IC). The acid halide and the baseare used in amounts of 1 to 3 equivalents, respectively, based on thehydroxy derivative. The reaction is carried out at 0° to 30° C. and iscompleted in 0.5 to 5 hours.

Step 3-5

[In the formulae, at least one of R^(3a) and R^(4a) is OCONR⁷ R⁸(wherein, R⁷ and R⁸ have the same meanings as defined above.)]

The hydroxy derivative obtained as a reaction intermediate in theabove-mentioned step 3-4 is allowed to react with 4-nitrophenylchloroformate in the presence of triethylamine in a solvent such asdichloromethane to give 4-nitrophenyl carbonate. 4-Nitrophenylchloroformate and the base are used in amounts of 1 to 3 equivalents,respectively, based on the hydroxy derivative. The reaction is carriedout at 0° to 30° C. and is completed in 0.5 to 5 hours.

Then the obtained carbonate is allowed to react with Compound (IV) whichis represented by the following formula:

    R.sup.7 R.sup.8 NH                                         (IV)

(wherein R⁷ and R⁸ have the same meanings as defined above), in thepresence of a base such as triethylamine and diisopropylethylamine in asolvent such as chloroform to give Compound (IC). Compound (IV) and thebase are used in amounts of 1 to 10 equivalents, respectively, based onthe carbonate. The reaction is carried out at 0° to 50° C. and iscompleted in 1 to 24 hours.

Step 3-6

[In the formulae, at least one of R^(3a) and R^(4a) is long-chainedalkyl (wherein the long-chained alkyl has the same meanings as definedabove.)]

The hydroxy derivative obtained as a reaction intermediate in theabove-mentioned step 3-4 is allowed to react with alkyl halide (whereinthe halogen is chloride, bromide or iodide) in the presence of a basesuch as sodium hydride in a solvent such as DMF to give Compound (IC).Alkyl halide and the base are used in amounts of 1 to 5 equivalents,respectively, based on the hydroxy derivative. The reaction is carriedout at -23° to 30° C. and is completed in 1 to 8 hours.

Production process 4

Compounds having various substituents at R³ and/or R⁴ positions can alsobe produced according to the following steps. ##STR6## (Wherein R¹, R²,W¹ and W² have the same meanings as defined above, and the definitionsof R^(3b) and R^(4b), and R^(3c) and R^(4c) are given in the followingsteps.)

Step 4-1

(In the formulae, at least one of R^(3b) and R^(4b) is nitro, and atleast one of R^(3c) and R^(4c) is amino.)

Compound (ID) is reduced in a solvent such as DMF, in the presence of10% Pd/C under a hydrogen atmosphere to give Compound (IE). 10% Pd/C isused in an amount of 10-200% (by weight) based on Compound (ID). Thereaction is carried out at 0° to 50° C. and is completed in 1 to 5hours.

Step 4-2

[In the formulae, at least one of R^(3b) and R^(4b) is amino, and atleast one of R^(3c) and R^(4c) is NR^(5a) R^(6a) (wherein R^(5a) andR^(6a) are the same or different and represent lower alkyl having thesame definitions as above)].

Compound (ID) is allowed to react with Compound (V) which is representedby the following formula:

    R.sup.15 CHO                                               (V)

(wherein, R¹⁵ represent alkyl having the number of carbon atoms which isless by one than that of the lower alkyl in the above-defined R^(5a) andR^(6a)), or a dimer thereof, in the presence of a reducing agent such assodium cyanoborohydride in a solvent such as a mixture of THF with water(10/1) at pH 5-6 to give Compound (IE). Sodium cyanoborohydride,Compound (V) and the dimer thereof are used in amounts of 1 to 1.5equivalents, respectively, based on Compound (ID). The reaction iscarried out at -10° to 50° C. and is completed in 1 to 8 hours.

Step 4-3

[In the formulae, at least one of R^(3b) and R^(4b) is amino, and atleast one of R^(3c) and R^(4c) is NHR^(5b) (wherein R^(5b) is an aminoacid residue where hydroxy group in the carboxylic acid is removed fromthe amino acid)].

Compound (ID) is allowed to react with an amino acid in the presence ofa condensation agent such as1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide in a solvent such asdichloromethane to give Compound (IE). The condensing agent and theamino acid are used in amounts of 1 to 3 equivalents, respectively,based on Compound (ID). The reaction is carried out at -10° to 50° C.and is completed in 1 to 24 hours. The amino acid to be used is onewhose amino group is protected by benzyloxycarbonyl group,t-butyloxycarbonyl group and the like. After completion of the reaction,deprotection is carried out by a conventional process such as catalyticreduction, treatment with an acid and the like.

Step 4-4

[In the formulae, at least one R^(3b) and R^(4b) is amino, and at leastone R^(3c) and R^(4c) is NHCONHR^(7a) (wherein, R^(7a) is a lower alkylhaving the same meaning as defined above)].

Compound (ID) is allowed to react with Compound (VI) which isrepresented by the following formula:

    R.sup.7a NCO                                               (VI)

(wherein R^(7a) has the same meaning as defined above) in a solvent suchas dichloromethane to give Compound (IE). Compound (VI) is used in anamount of 1 to 10 equivalents based on Compound (ID). The reaction iscarried out at 0° to 100° C. and is completed in 1 to 5 hours.

Step 4-5

[In the formulae, at least one of R^(3b) and R^(4b) is amino, and atleast one of R^(3c) and R^(4c) is NHCO(CH₂)_(m) NR⁷ R⁸ (wherein R⁷, R⁸and m have the same meanings as defined above.)]

Compound (ID) is allowed to react with Compound (VII) which isrepresented by the following formula (VII):

    Hal.sup.1 -(CH.sub.2).sub.m CO--Hal.sup.2                  (VII)

(wherein Hal¹ and Hal² are the same or different, and have the samemeanings as defined for the above-mentioned Hal, and m has the samemeaning as defined above) in the presence of a base such astriethylamine in a solvent such as dichloromethane to give amidederivative. Compound (VII) and the base are used in amounts of 1 to 3equivalents, respectively, based on Compound (ID). The reaction iscarried out at 0° to 30° C. and is completed in 0.5 to 5 hours.

Then the amide is allowed to react with Compound (IV) in the presence ofa base such as triethylamine and diisopropylethylamine in a solvent suchas chloroform to give Compound (IE). Compound (IV) and the base are usedin amounts of 1-10 equivalents, respectively, based on the amidederivative. The reaction is carried out at 20° to 100° C. and iscompleted in 1 to 24 hours.

Step 4-6

[In the formulae, at least one of R^(3b) and R^(4b) is hydroxymethyl,and at least one of R^(3c) and R^(4c) is CH₂ OR^(12a) (wherein R^(12a)represents all the groups but hydrogen in the definition of R¹²) or CH₂SR^(13a) (wherein R^(13a) represents all the groups but hydrogen in thedefinition of R¹³)].

Compound (ID) is allowed to react with Compound (VIII) or Compound (IX)which is represented by the following formula (VIII) or (IX);

    R.sup.12a OH                                               (VIII)

or

    R.sup.13a SH                                               (IX),

(wherein R¹² and R¹³ have the same meanings defined as above)

in the presence of camphorsulfonic acid in a solvent such as chloroformto give Compound (IE). Camphorsulfonic acid is used in an amount of 1 to20 equivalents, and Compound (VIII) or Compound (IX) is used in anamount of 5 to 20 equivalents, respectively, based on Compound (ID). Thereaction is carried out at 0° to 50° C. and is completed in 1 to 24hours.

Step 4-7

(In the formulae, at least one of R^(3b) and R^(4b) is substituted orunsubstituted lower alkenyl, and at least one of R^(3c) and R^(4c) issubstituted or unsubstituted lower alkyl.)

Compound (ID) is hydrogenated in a solvent such as a mixture of DMF andacetic acid (2/1), in the presence of 20% Pd(OH)₂ /C under a hydrogenatmosphere, to give Compound (IE). 20% Pd(OH)₂ /C is used in an amountof 10-200% (by weight) based on Compound (ID). The reaction is carriedout at 0° to 50° C. and is completed in 1 to 5 hours.

Step 4-8

(In the formulae, at least one of R^(3b) and R^(4b) is lower alkylsubstituted with hydroxy, and at least one of R^(3c) and R^(4c) is loweralkyl.)

Compound (ID) is allowed to react with triethylsilane in the presence oftrifluoroacetic acid in a solvent such as dichloromethane to giveCompound (IE). Trifluoroacetic acid is used in an amount of 10 to 100equivalents and triethylsilane is used in an amount of 3 to 10equivalents, based on Compound (ID). The reaction is carried out at 0°to 50° C. and is completed in 10 minutes to 8 hours.

Production process 5

Compound (IG) [Compound (I) in which R¹ is hydroxy-substituted loweralkyl]can be produced not only according to the Production process 1 butalso according to the following steps. ##STR7## (wherein, R², R³, R⁴, W¹and W² have the same meanings as defined above, and the definitions ofR^(1b) and R^(1c) are given in the following steps.)

Step 5-1

[In the formulae, R^(1b) is --(CH₂)_(n) CH═CH₂ (wherein n is an integerof 0-4), and R^(1c) is --(CH₂)_(n) CH(OH)CH₂ OH (wherein n has the samemeaning as defined above].

Compound (IF) obtained according to a process similar to Productionprocess 1 is allowed to react with osmium tetroxide in a solvent such asa mixture of chloroform and acetone (1/1) in the presence ofN-methylmorpholine N-oxide to give Compound (IG). Osmium tetroxide isused in an amount of 0.02 to 1 equivalent, and N-methyl morpholineN-oxide is used in an amount of 1 to 5 equivalents based on Compound(IF). The reaction is carried out at -10° to 50° C. and is completed in1 to 8 hours.

Step 5-2

[In the formulae, R^(1b) is --(CH₂)_(n) CH═CH₂ (wherein n has the samemeaning as defined above) and R^(1c) is --(CH₂)_(n) CH₂ CH₂ OH or--(CH₂)_(n) CH(OH)CH₃ (wherein n has the same meaning defined asabove)].

Compound (IF) is allowed to react with a reducing agent such asborane-THF complex in a solvent such as THF, and treated with 30%aqueous hydrogen peroxide in the presence of 3M sodium hydroxide to giveCompound (IG). The borane-THF complex is used in an amount of 0.5 to 1.5equivalents based on Compound (IF) and 3M sodium hydroxide and 30%aqueous hydrogen peroxide are used in amounts of 0.1 to 1 ml,respectively, based on 1 mmol of Compound (IF). The reaction is carriedout at -10° to 50° C. and is completed in 1 to 8 hours.

Step 5-3

[In the formulae, R^(1b) is --(CH₂)_(n) CH(OH)CH₂ OH (wherein n has thesame meaning as defined above) and R^(1c) is --(CH₂)_(n) CH₂ OH (whereinn has the same meaning as defined above)].

Compound (IF) is allowed to react with an oxidizing agent such as sodiumperiodate in a solvent such as a mixture of THE and water (5/3) to giveformyl derivative wherein R^(1b) is converted to --(CH₂)_(n) CHO. Theobtained formyl derivative is allowed to react with a reducing agentsuch as sodium borohydride in an appropriate solvent such as a mixtureof THF and methanol (1/1) to give Compound (IG). The sodium periodate isused in an amount of 1 to 3 equivalents based on Compound (IF). Thereaction is carried out at -10° to 50° C. and is completed in 1 to 8hours.

A functional group contained in R¹, R², R³ or R⁴ can be also changed byother known processes in addition to the above processes [for example,R. C. Larock, Comprehensive Organic Transformations (1989)].

Isolation and purification of the products in the above processes can becarried out using an appropriate combination of methods conventionallyused in organic synthesis, for example, filtration, extraction, washing,drying, concentration, crystallization and various types ofchromatography. The reaction intermediate can be used in the subsequentreaction without purification.

In the case where a salt of Compound (I) is desired, and it is producedin the form of the desired salt, it can be subjected to purification assuch. In the case where Compound (I) is produced in the free state andits salt is desired, Compound (I) is dissolved or suspended in asuitable solvent, followed by addition of an acid to form a salt.

Compound (I) or a pharmaceutically acceptable salt thereof may alsoexist in the form of an adduct with water or various solvents. Theseadducts are within the scope of the present invention.

Examples of Compound (I) are listed in Table 1.

                                      TABLE 1                                     __________________________________________________________________________     ##STR8##                                                                     Compd.                                                                              W.sup.1,W.sup.2                                                                     R.sup.1    R.sup.2                                                                          R.sup.3         R.sup.4         Salt                __________________________________________________________________________    1     H,H   CH.sub.2 OH                                                                              H  H               H                                   2     H,H   CH.sub.2 CH(OH)CH.sub.2 OH                                                               H  H               H               HCl                 3     H,H   (CH.sub.2).sub.2 OH                                                                      H  H               H                                   4     H,H   (CH.sub.2).sub.3 OH                                                                      H  H               H                                   5     H,H   CH.sub.2 CH(OH)CH.sub.3                                                                  H  H               H                                   6     H,H   Me         H  CH.sub.2 OH     H                                   7     H,H   Me         H  CH.sub.2 OH     CH.sub.2 OH                         8     H,H   Me         H  Me              H                                   9     H,H   Me         H  Me              Me              HCl                 10    H,H   Me         H  Et              Et              HCl                 11    H,H   Me         H                                                                                 ##STR9##       H                                   12    H,H   Me         H  NH.sub.2        H               2HCl                13    H,H   Me         H  NMe.sub.2       H               2HCl                14    H,H   Me         H                                                                                 ##STR10##      H               2HCl                15    H,H   Me         H                                                                                 ##STR11##      H               3HCl                16    H,H   Me         H  NH.sub.2        NH.sub.2        3HCl                17    H,H   Me         H  NMe.sub.2       NMe.sub.2       3HCl                18    H,H   Me         H                                                                                 ##STR12##                                                                                     ##STR13##      3HCl                19    H,H   Me         H                                                                                 ##STR14##                                                                                     ##STR15##      HCl                 20    H,H   Me         H                                                                                 ##STR16##                                                                                     ##STR17##      5HCl                21    H,H   Me         H  OAc             OAc             HCl                 22    H,H   Me         H  OCO.sup.t Bu    OCO.sup.t Bu    HCl                 23    H,H   Me         H  OCO(CH.sub.2).sub.14 Me                                                                       OCO(CH.sub.2).sub.14 Me                                                                       HCl                 24    H,H   Me         H                                                                                 ##STR18##                                                                                     ##STR19##      HCl                 25    H,H   Me         H                                                                                 ##STR20##                                                                                     ##STR21##      3HCl                26    H,H   Me         H  CH.sub.2 CH.sub.2 CO.sub.2 Et                                                                 H               AcOH                27    H,H   Me         H                                                                                 ##STR22##      H               3HCl                28    H,H   Me         H  CNNMe.sub.2     H                                   29    H,H   Me         H                                                                                 ##STR23##      H               2HCl                30    H,H   Me         H  CH.sub.2 S(CH.sub.2).sub.2 NMe.sub.2                                                          H                                   31    H,H   Me         H  CH.sub.2 O(CH.sub.2).sub.2 NMe.sub.2                                                          H                                   32    H,H   Me         H  CH.sub.2 SEt    CH.sub.2 SEt                        33    H,H   Me         H  CH.sub.2 S(CH.sub.2).sub.2 NMe.sub.2                                                          CH.sub.2 OH                         34    O     Me         H  CH.sub.2 OH     CH.sub.2 OH                         35    H,H   Me         H  OCO(CH.sub.2).sub.7 Me                                                                        OCO(CH.sub.2).sub.7 Me                                                                        HCl                 36    H,H   Me         H  OCOCH.sub.2 (OCH.sub.2 CH.sub.2).sub.3 OMe                                                    OCOCH.sub.2 (OCH.sub.2 CH.sub.2)                                              .sub.3 OMe                          37    H,H   Me         H  NHCO(CH.sub.2).sub.14 Me                                                                      NHCO(CH.sub.2).sub.14 Me                                                                      HCl                 38    H,H   Me         H  NHCO(CH.sub.2).sub.7 Me                                                                       NHCO(CH.sub.2).sub.7 Me                                                                       HCl                 39    H,H   Me         H  O(CH.sub.2).sub.15 Me                                                                         O(CH.sub.2).sub.15 Me                                                                         HCl                 40    H,H   Me         H  O(CH.sub.2).sub.8 Me                                                                          O(CH.sub.2).sub.8 Me                                                                          HCl                 __________________________________________________________________________

Compound (I) and pharmaceutically acceptable salts thereof can be usedas such or in the form of various pharmaceutical compositions accordingto their pharmacological activity and the intended administrationpurpose. The pharmaceutical compositions according to the presentinvention can be prepared by uniformly mixing an effective amount ofCompound (I) or a pharmaceutically acceptable salt thereof as an activeingredient with pharmaceutically acceptable carriers. The carriers mayhave a wide range form depending on the type of the preparation desiredfor the administration. The pharmaceutical compositions are preferablyformulated into a unit dose form which is suited to oral or non-oraladministration. The dose forms for non-oral administration includeointments and injections.

Tablets can be prepared using, in a conventional manner, excipients suchas lactose, glucose, sucrose, mannitol, and methyl cellulose;disintegrating agents such as starch, sodium alginate, calciumcarboxymethyl cellulose, and crystalline cellulose; lubricants such asmagnesium stearate and talc; binders such as gelatin, polyvinyl alcohol,polyvinylpyrrolidone, hydroxypropyl cellulose, and methyl cellulose;surface active agents such as sucrose fatty acid esters and sorbitolfatty acid esters; and the like. Tablets each containing 50 to 200 mg ofan active ingredient are appropriate.

Granules can be prepared using, in a conventional manner, excipientssuch as lactose and sucrose; disintegrating agents such as starch;binders such as gelatin; and the like. Powders are prepared usingexcipients such as lactose and mannitol, and the like in a conventionalmanner. Capsules are prepared using, in a conventional manner, gelatin,water, sucrose, gum arabic, sorbitol, glycerin, crystalline cellulose,magnesium stearate, talc, etc. Capsules each containing 50 to 200 mg ofan active ingredient are appropriate. Syrups are prepared usingsaccharides such as sucrose, water, ethanol, etc. in a conventionalmanner.

For the preparation of ointments, ointment bases such as vaseline,liquid paraffin, lanolin, and macrogol, and emulsifying agents such assodium lauryl lactate, benzalkonium chloride, sorbitan monofatty acidesters, sodium carboxymethyl cellulose, and gum arabic, and the like maybe used in a conventional manner.

Injectable preparations can be prepared using, in a conventional manner,solvents such as water, physiological saline, vegetable oil (e.g., oliveoil and peanut oil), ethyl oleate, and propylene glycol; solubilizingagents such as sodium benzoate, sodium salicylate, and urethane;isotonizing agents such as sodium chloride and glucose; preservativessuch as phenol, cresol, p-hydroxybenzoic esters, and chlorobutanol;antioxidants such as ascorbic acid and sodium pyrosulfite; and the like.

Compound (I) and pharmaceutically acceptable salts thereof may beadministered orally or non-orally as an ointment or an injection. Theeffective dose and the administration schedule vary depending on theadministration route, the age, body weight and symptoms of the patient,and the like, but generally ranges 6.0 to 300 mg/kg/day in a single to 4divided doses.

The toxicity and pharmacological activity of Compound (I) will bedescribed by way of Test Examples.

TEST EXAMPLE 1 Megakaryocyte colony formation-stimulating activity

An eight-weeks-old BALB/c mouse was killed. Its femurs and cervicalvertebrae were taken out, and both end sections thereof were cut off.Bone marrow cells were collected from the pieces cut off from the femursand cervical vertebrae using a syringe containing IMDM (430-2200EAprepared by Gibco Co.), and then blown into a test tube. The test tubewas allowed to stand for 5 minutes, and the supernatant was collectedwith a piper. To a reaction mixture comprising the bone marrow cells(50,000 cells), bovine serum albumin (2%: A4508 made by Sigma Co.),transferrin (600 μg/ml: 652202 made by Boehringer Mannheim Co.), IL-3(100 U/ml), cholesterol (16 μg/ml: 036-0641 made by Wako Co.) and agar(0.6%: 0142-02 made by Difco Laboratories) were separately added thetest compounds at various concentrations, and 1 ml each of the mixtureswas put into a 35-mm dish (Lux Co.), followed by incubation under theconditions of 37° C. 5% CO₂ and a humidity of 95% or more for 7 days.Separately, IL-3 alone was added to the bone marrow cells to prepare acontrol. After the incubation was completed, the agar was dried over afilter paper (1001-055 made by Whatman Co.) and then fixed with 2.5%glutaraldehyde, followed by acetylcholinesterase staining (ACHEstaining).

The ACHE staining was carried out by the method described below.

ACHE staining: To each sample was added a solution comprising 0.67 mg/mlacetylthiocholine iodide, 2.94 mg/ml sodium citrate, 7.5 mg/ml copper(II) sulfate and 1.65 mg/ml potassium ferricyanide, and the mixture wasallowed to stand at room temperature in the dark for 4-6 hours.

A group of 4 or more megakaryocytes which were stained reddish brown wasregarded as a colony, and the number of colonies per dish was calculatedusing a microscope. The results are shown in Table 2 as relative valuesto the control.

(The table shows the relative values calculated on the basis of thecontrol defined as 100.)

                  TABLE 2                                                         ______________________________________                                        Compd.        Concn.(nM) Rel. Value                                           ______________________________________                                        Control       --         100                                                   1            1          129                                                   8            10         157                                                  12            10         126                                                  16            1          125                                                  28            1          134                                                  30            1          116                                                  ______________________________________                                    

TEST EXAMPLE 2 Platelet Production-Stimulating Activity in Mice

A test compound was intraperitoneally administered to four 7-week-oldmale BALB/c mice per group once a day for consecutive 5 days (day 1 today 5). A control group (4 mice per group) received only the solvent (5%Tween water). The blood was collected from the fundus oculi vein of eachanimal on the 15th day from the start of administration (day 15), andthe number of the platelets was counted with a microcell counter (ModelCC-180A, manufactured by Toa Iryo Denshi Co.). The rate of increase ofthe number of platelets in the test group (average) over the control(average) was calculated according to the following formula to evaluatethe effect of the test compound. The results obtained are shown in Table3.

                  TABLE 3                                                         ______________________________________                                        Rate of Increase = A/B × 100                                            A: the number of platelets in test group                                      B: the number of platelets in control group                                   Test Compd. Dose (mg/kg)                                                                              Rate of increase (%)                                  ______________________________________                                         8          25          134                                                   22          50          168                                                   28          12.5        156                                                   ______________________________________                                    

TEST EXAMPLE 3 Acute Toxicity

A solution (0.2 ml) of a test compound in phosphate-bufferedphysiological saline was intraperitoneally administered to a 6-week-oldmale DDY mice (3 mice per group). The 50% lethal dose (LD₅₀) wascalculated from the survival rate after 24 hours from theadministration. As a result, all the Compounds 1 to 40 tested had anLD₅₀ of not less than 10 mg/kg.

Certain embodiments of the present invention are illustrated in thefollowing representative examples. In the following description, brine,Na₂ SO₄, NaHCO₃, CH₂ Cl₂, CHCl₃, MeOH, AcOEt, Cbz, and TLC, respectivelystand for a saturated aqueous solution of sodium chloride solution,sodium sulfate, sodium hydrogen carbonate, dichloromethane, chloroform,methanol, ethyl acetate, benzyloxycarbonyl, and thin layerchromatography. In the following description, the position of thesubstituent which is bonded in the staurosporine skeleton is defined asfollows; R¹ is bonded to a nitrogen at the 2-position, R² is bonded to anitrogen at the 11-position, R³ is bonded to a carbon at the17-position, R⁴ is bonded to a carbon at the 5-position, and W is bondedto a carbon at the 3-position. The staurosporine derivative, as thestarting material, was prepared from staurosporine by such knownprocesses as those given in WO89/07105, WO88/07045 and the like, or by aprocess analogous to those processes.

Example 1

Compound 1

To 11 mg (0.02 mmol) of 2-formylstaurosporine dissolved in 0.2 ml of THFand 0.2 ml of MeOH was added 4 mg (0.06 mmol) of sodium borohydride,followed by stirring at a room temperature for 1.5 hours. The reactionmixture was diluted with CHCl₃, then washed with water and brine, andthen dried over Na₂ SO₄. The solvent was removed by evaporation, and theresidue was purified by preparative TLC (MeOH/CHCl₃ 5/95) to give 5 mg(51%) of Compound 1.

¹ HNMR(DMSO-d₆) δ;2.335(s,3H), 4.109(br.s,1H), 5.120(s,2H),5.971(t,1H,J=6.9 Hz), 6.737 (br.s, 1H), 7.262-8.019 (m,7H),9.290(d,1H,J=8.1 Hz).

Fab-MS(m/z); 497(M+1)⁺

Example 2

Compound 2

To 1.8 g (3 mmol) of 11-Cbz-staurosporine dissolved in 30 ml of DMF wasadded 180 mg (4.5 mmol) of 60% sodium hydride at 0° C. followed bystirring for 10 minutes Then, 0.39 ml (4.5 mmol) of allyl bromide wasadded thereto at the same temperature, followed by stirring for 2.5hours. The reaction mixture was diluted with CHCl₃, washed with waterand brine, and then dried over Na₂ SO₄. The solvent was removed byevaporation, and the residue was purified by silica gel columnchromatography (AcOEt/toluene 1/9) to give 1.17 g (61%) of 2-allylderivative.

¹ HNMR (DMSO-d₆, 90° C.) δ; 2.308(s,3H), 2.646(s,3H), 2.748(s,3H),4.221(br.s,1H), 4.341(br.d,2H,J=5.6 Hz), 4.683(m,1H), 5.032(s,2H),5.165-5.351(m,4H), 6.007-6.090(m,1H), 6.958(dd,1H,J=6.1,8.8 Hz),7.259-7.512(m,9H), 8.039(d,1H,J=7.8 Hz), 9.310(d,1H,J=8.1 Hz).

Fab-MS(m/z); 641(M+1)⁺

422 mg (0.66 mmol) of the 2-allyl derivative was dissolved in a mixtureof 5 ml of CHCl₃ and 5 ml of acetone. A solution of 8.3 mg (0.033 mmol)of osmium tetroxide in 0.083 ml of acetone and 116 mg (0.99 mmol) ofN-methylmorpholine oxide were added thereto, followed by stirring atroom temperature for 1.5 hours. 10 ml of 8% sodium thiosulfate solutionwas added thereto and the mixture was stirred for 1 hour. Chloroform wasadded to the reaction mixture and the organic layer was separated,washed with brine and dried over Na₂ SO₄. The solvent was removed byevaporation. The residue was purified by silica gel columnchromatography (MeOH/CHCl₃ 5 /95) to give 422 mg (95%) of 11-Cbzderivative of Compound 2.

¹ HNMR(DMSO-d₆,90° C.) δ;2.312(s,3H), 2.636(s,3H), 2.746(s,3H),3.489(d,2H,J=5.6 Hz), 3.686(m,1H), 3.850(m,1H), 3.971(m,1H),4.220(br.s,1H), 4.684(m,1H),5.116-5.253(m,4H), 6.958(dd,1H,J=5.8,8.8Hz), 7.256-7.513(m,9H), 7.583(d,1H,J=8.6 Hz), 8.032(d,1H,J=7.3 Hz),9.315(d,1H,J=7.8 Hz).

Fab-MS(m/z); 675(M+1)⁺

To a solution of 30 mg (0.044 mmol) of the 11-Cbz derivative of Compound2 dissolved in 1 ml of DMF was added 30 mg of 10% Pd/C, followed bystirring for 2 hours at 50° C. under a hydrogen atmosphere. The reactionmixture was filtered with celite and the solvent was removed byevaporation. The residue was purified by preparative TLC (MeOH/CHCl₃2/8), and then treated with 0.6N HCl/AcOEt to give 5.5 mg (21%) ofhydrochloride of Compound 2.

¹ HNMR(DMSO-d₆,90° C.) δ; 2.191(m,1H), 2.417(s,3H), 2.675(s,3H),3.504(d,2H,J=5.5 Hz), 3.686(m,1H), 3.664-3.731(m,1H), 3.825-3.883(m,1H),3.975(m,2H), 4.520(s,1H), 5.126-5.223(m,2H), 6.903(dd,1H,J=3.6,9.3 Hz),7.287-8.075(m,7H), 9.355(d,1H,J=7.9 Hz).

Fab-MS(m/z); 541(M+1)⁺

Example 3

Compound 3

To a solution of 25 mg (0.037 mmol) of the 11-Cbz derivative of Compound2 dissolved in 0.5 ml of THF was added under cooling with ice 18 mg(0.078 mmol) of sodium periodate dissolved in 0.3 ml of water, followedby stirring at room temperature for 3.5 hours. The reaction mixture wasdiluted with CHCl₃, washed with 5% aqueous sodium thiosulfate and brine,and then dried over Na₂ SO₄. The solvent was removed by evaporation, andthe residue was purified by preparative TLC (MeOH/CHCl₃ 5 /95) to give19 mg (yield: 80%) of 2-(2-oxoethyl) derivative.

¹ HNMR(DMSO-d₆,90° C.) δ; 2.321(s,3H), 2.641(s,3H), 2.753(s,3H),4.222(t,1H,J=1.8 Hz), 4.584(s,2H), 4.694(m,1H), 5.141(s,2H),5.184(d,1H,J=12.5 Hz), 5.238(d,2H,J=12.4 Hz), 6.958(dd,1H,J=5.9,8.8 Hz),7.259-8.026(m,12H), 9.257(d,1H,J=7.8Hz), 9.792(s,1H).

Fab-MS(m/z); 643(M+1)⁺

11-Cbz derivative of Compound 3 was obtained from 64 mg (0.1 mmol) ofthe 2-(2-oxoethyl) derivative by the procedure analogous to that ofExample 1 (yield:49 mg, 77%).

¹ HNMR(DMSO-d₆, 90° C.) δ;2.658(br.s,3H), 2.740(s,3H), 4.270(br.s,1H),4.691(m,1H), 4.895(t,1H,J=4.5 Hz), 5.160(s,2H), 5.198(s,2H),7.010(br.s,1H), 7.276-8.073(m,12H), 9.311(d,1H,J=7.8 Hz).

Fab-MS(m/z); 645(M+1)⁺

Compound 3 was obtained from 25 mg (0.038 mmol) of the 11-Cbz derivativeof Compound 3 by the procedure analogous to that of Example 2 (yield: 17mg, 83%).

¹ HNMR(DMSO-d₆) δ;1.159(s,3H), 2.322(s,3H), 3.300(s,3H),3.737-3.827(m,4H), 4.061(d,1H,J=3.4 Hz), 4.614(t,1H,J=4.9 Hz),5.092(s,2H), 6.686(dd,1H,J=2.2,5.9 Hz), 7.227-7.984(m,7H),9.313(d,1H,J=7.6 Hz).

Fab-MS(m/z); 511(M+1)⁺

Example 4

Compound 4 and Compound 5

To a solution of 128 mg (0.2 mmol) of the 2-allyl-11-Cbz-staurosporinein 2 ml of THF was added 0.2 ml (0.2 mmol) of 1M borane-THF complex,followd by stirring at room temperature for 1 hour. To the reactionmixture were added 0.06 ml of water, 0.08 μl of 3M aqueous NaOH and 0.08ml of 30% aqueous hydrogen peroxide, and the mixture was stirred at roomtemperature for 1 hour. The reaction mixture was diluted with CHCl₃,washed with water and brine, and then dried over Na₂ SO₄. After thesolvent was removed by evaporation, the residue was purified bypreparative TLC (AcOEt/toluene 7/3) to give 40 mg (31%) of 11-Cbzderivative of Compound 4 and 37 mg (28%) of 11-Cbz derivative ofCompound 5.

11-Cbz derivative of Compound 4

¹ HNMR(DMSO-d₆) δ; 1.936(quint, 2H,J=6.6 Hz), 2.670(br.s,3H),2.738(s,3H), 3.555(t,2H,J=6.2 Hz), 3.715-3.830(m,2H), 4.271(br.s,1H),5.099(s,2H), 5.195(s,2H), 7.000(br.s,1H), 7.271-8.079(m,11H),8.809(d,1H,J=7.5 Hz), 9.304(dd,1H,J=1.0,8.0 Hz).

Fab-MS(m/z); 659(M+1)⁺

11-Cbz derivative of Compound 5

¹ HNMR(DMSO-d₆) δ;1.178(d,3H,J=6.1 Hz), 2.664(br.s,3H), 2.738(s,3H),3.579-3.713(m,2H), 4.124(m,1H), 4.268(br.s,1H), 4.692(br.s,1H),4.918(br.s,1H), 5.122-5.231(m,4H), 7.002(br.s,1H), 7.271-8.053(m,12H),9.312(d,1H,J=7.8 Hz).

Fab-MS(m/z); 659(M+1)⁺

Compound 4 and Compound 5 were obtained from 28 mg (0.042 mmol) of the11-Cbz derivative of Compound 4 and 25 mg (0.038 mmol) of the 11-Cbzderivative of Compound 5, respectively by the procedure analogous tothat of Example 2 (yield of Compound 4: 13 mg, 59%; yield of Compound 5:15 mg, 75%).

Compound 4

¹ HNMR(DMSO-d₆) δ; 1.477(br.s,3H), 1.931(quint,2H,J=6.5 Hz),2.314(s,3H), 3.555(q,2H,J=6.1 Hz), 4.045(d,1H), 4.569(t,1H,J=5.2 Hz),5.053(s,2H), 6.718(t,1H,J=3.5 Hz), 7.246-7.998(m,7H), 9.287(d,1H,J=7.4Hz).

Fab-MS(m/z); 525(M+1)⁺

Compound 5

¹ HNMR(DMSO-d₆) δ;1.180(d,3H,J=6.1 Hz), 1.476(br.s,3H), 2.314(s,3H),4.083(d,1H,J=3.5 Hz), 4.108(m,1H), 4.912(dd,1H,J=4.0,4.6 Hz),5.084-5.177(m,2H), 6.718(t,1H,J=3.7 Hz), 7.244-8.000(m,7H),9.294(dd,1H,J=0.9,7.9 Hz).

Fab-MS(m/z); 525(M+1)⁺

Example 5

Compound 6 and Compound 7

To a solution of 480 mg (1 mmol) of 2-methyl staurosporine in 20 ml ofdichloroethane was added 0.28 ml (3 mmol) of dichloromethyl methyl etherand 0.32 ml (3 mmol) of titanium tetrachloride, followed by stirring atroom temperature for 2 hours. The reaction mixture was diluted withCHCl₃ and poured to a saturated aqueous solution of NaHCO₃, and theresulting organic layer was separated, washed with brine, and then driedover Na₂ SO₄. The solvent was removed by evaporation and the residue waspurified by silica gel column chromatography (MeOH/CHCl₃ 2/98) to give378 mg of a mixture of 17-formyl derivative and 5,17-diformylderivative.

Compound 6 and Compound 7 were obtained from 51 mg (0.1 mmol) of theresulting mixture by the procedure analogous to that of Example 1 (yieldof Compound 6: 44 mg, 86% yield of Compound 7: 7 mg, 13%).

Compound 6

¹ HNMR(DMSO-d₆) δ;1.440(br.s,3H), 2.310(s,3H), 4.073(d,1H,J=3.4 Hz),4.676(d,2H,J=5.7 Hz), 5.025(s,2H), 5.116(t,1H,J=5.8 Hz),6.995(t,1H,J=3.6 Hz), 7.265-7.992(m,7H), 9.232(d,1H,J=1.0 Hz).

Fab-MS(m/z); 511(M+1)⁺

Compound 7

¹ HNMR(DMSO-d₆) δ;4.081(br.s,1H), 4.683(t,4H,J=6.0 Hz), 5.020(s,2H),5.118(t,1H,J=5.6 Hz), 5.116(t,1H,J=5.7 Hz), 6.704(br.s,1H),7.371-7.940(m,5H), 9.228(d,1H,J=1.0 Hz).

Fab-MS(m/z); 541(M+1)⁺

Example 6

Compound 8

To a solution of 102 mg (0.2 mmol) of Compound 6 in 2 ml of acetic acidwere added 0.077 ml (1 mmol) of trifluoroacetic acid and 0.096 ml (0.6mmol) of triethylsilane, followed by stirring at room temperature for5.5 hours. The reaction mixture was diluted with CHCl₃, poured to asaturated aqueous solution of NaHCO₃, and then the organic layer wasseparated and washed with brine, and dried over potassium carbonate (K₂CO₃). After the solvent was removed by evaporation, the residue waspurified by preparative TLC (MeOH/CHCl₃ 5/95) to give 50 mg (51%) ofCompound 8.

¹ HNMR(DMSO-d₆) δ;1.494(br.s,3H), 2.310(s,3H), 2.520(s,3H), 3.257(s,3H),4.081(br.s,1H), 5.018(s,2H), 6.675(br.t,1H,J=3.6 Hz), 7.270-7.988(m,6H),9.105(t,1H,J=0.7 Hz).

Fab-MS(m/z); 495(M+1)⁺

Example 7

Compound 9

To a solution of 99 mg (0.185 mmol) of Compound 7 in 1 ml ofdichloromethane were added 0.18 ml(1.1 mmol) of triethylsilane and 1.0ml (13 mmol) of trifluoroacetic acid in an argon atmosphere, followd bystirring overnight at room temperature. To the reaction mixture wereadded ice and an aqueous sodium hydroxide to stop the reaction, followedby extraction with dichloromethane. The extract was washed with brineand dried over MgSO₄, and the solvent was removed by evaporation. Theresidue was purified by TLC (CHCl₃ /MeOH/triethylamine 25/1/1) anddissolved in a mixed solvent of 1 ml of chloroform and 2 ml of AcOEt.0.10 ml of 0.88N HCl (dissolved in AcOEt) was added to the solution andthe mixture was stirred at room temperature for 1 hour. The resultingprecipitate was collected by filtration, and washed with AcOEt and driedunder reduced pressure to give 12 mg (19%) of Compound 9.

¹ HNMR(DMSO-d₆) δ;2.0-2.2(m, 1H), 2.267(s,3H), 2.465 (s,3H),2.527(s,3H), 2.548(s,3H), 2.688(br.s,3H), 3.271(s,3H), 3.9-4.1(br.s,1H),4.398(br.s,1H), 5.072(s,2H), 6.87-6.90 (m,1H), 7.32-7.37(m,2H),7.453(d,1H,J=8.1 Hz), 7.877(s,1H), 7.949(d,1H,J=8.8 Hz),8.5-8.7(br.s,2H), 9.139(d,1H,J=0.7 Hz).

Fab-MS(m/z); 509(M+1)⁺

Example 8

Compound 10

A solution of 5.43 g (40.7 mmol) of aluminum chloride in 200 ml of CH₂Cl₂ was cooled to 0° C., and 5.76 ml (81.4 mmol) of acetyl chloride wasadded thereto, followed by stirring for 30 minutes. The mixture wascooled to -78° C. A solution of 5.00 g (8.14 mmol) of2-methyl-11-Cbz-staurosporine in 50 ml of CH₂ Cl₂ was added dropwise tothe mixture, followed by stirring for 1 hour. The reaction was stoppedby the addition of a saturated aqueous solution of sodium hydrogencarbonate, followed by extraction with CHCl₃. The extract was washedwith water and brine, and then dried over Na₂ SO₄. The solvent wasremoved by evaporation and the residue was purified by silica gel columnchromatography (MeOH/CHCl₃ 1/200) to give 4.73 g (83% of 5,17-diacetylderivative.

¹ HNMR(DMSO-d₆) δ; 2.690(s,6H), 2.738(s,3H), 2.755(s,3H), 5.234(s,4H),7.077(br.s,1H), 7.402(br.s,5H), 7.724(d,1H,J=8.6 Hz), 8.086-8.106(m,3H),8.587(s,1H), 10.032(s,1H)

Fab-MS(m/z); 565(M+1)⁺

A solution of 159 mg (0.23 mmol) of the 5,17-diacetyl derivative in 10ml of CH₂ Cl₂ and 5 ml of MeOH was cooled to 0° C., and 26 mg (0.68mmol) of sodium borohydride was added thereto, followed by stirring for3.5 hours. After the completion of reaction, the mixture was dilutedwith CHCl₃, washed with brine, and then dried over Na₂ SO₄. After thesolvent was removed by evaporation, the residue was purified by silicagel column chromatography (MeOH/CHCl₃ 1/20) to give 140 mg (87%) of5,17-bis(2-hydroxyethyl) derivative.

¹ HNMR(DMSO-d₆,90° C.) δ; 1.429-1.479(m,6H), 2.288(s,3H), 2.636(s,3H),2.744(s,3H), 3.289(s,3H), 4.191(s,1H), 4.688(d,1H,J=11.0 Hz),4.917-4.997(m,2H), 5.041(s,2H), 5.193(d,1H,J=12.5 Hz), 5.251(d,1H,J=12.5Hz), 6.923(dd,1H,J=5.9,8.0 Hz), 7.516-7.348(m,8H), 7.833(dd,1H,J=3.4,8.8Hz), 7.999(dd,1H,J=1.7,12.2 Hz), 9.297(d,1H,J=9.0 Hz)

Fab-MS(m/z);702(M)⁺

To a solution of 140 mg (0.20 mmol) of the 5,17-bis(2-hydroxyethyl)derivative in 5 ml of CH₂ Cl₂ was added 191 μl (1.20 mmol) oftriethylsilane and 1.53 ml (19.9 mmol) of trifluoroacetic acid, followedby stirring at room temperature for 10 minutes. The mixture was madebasic with 1.0N NaOH, followed by extraction with CHCl₃, and the extractwas washed with brine and dried over Na₂ SO₄. The solvent was removed byevaporation and the residue was purified by preparative TLC (MeOH/CHCl₃3/100) to give 107 mg (80%) of 11-Cbz derivative of Compound 10. ¹HNMR(DMSO-d₆, 90° C.) δ; 1.302-1.364(m,6H), 2.275(s,3H), 2.625(s,3H),2.740(s,3H), 2.793-2.876(m,4H), 3.278(s,3H), 4.173(s,1H),4.640-4.690(m,1H), 5.039(s,2H), 5.190(d,1H,J=12.5 Hz), 5.248(d,1H,J=12.5Hz), 6.909(dd,1H,J=5.8,8.8 Hz), 7.301-7.478(m,8H), 7.794(d,1H,J=8.6 Hz),7.827(s,1H), 9.175(s,1H)

Fab-MS(m/z); 671(M+1)⁺

To a solution of 106 mg (0.16 mmol) of the 11-Cbz derivative of Compound10 in 10 ml of DMF was added 100 mg of 10% Pd/C, followed by stirringunder a hydrogen atmosphere at room temperature for 3.5 hours. After thecompletion of reaction, the reaction mixture was filtered with celiteand the solvent was removed by evaporation. The residue was dissolved in5 ml of CHCl₃ and ml of 0.88N HCl/AcOEt was added thereto, followed bystirring for 1 hour. The resulting precipitate was filtered off to give72 mg (79%) of Compound 10. ¹ HNMR(DMSO-d₆,90° C.) δ; 1.306-1.408(m,6H),2.137-2.206(m,1H), 2.444(s,3H), 2.583(br.s,3H), 2.802-2.885(m,4H),3.281(s,3H), 3.885(br.s,1H), 4.462(s,1H), 5.046(s,2H),6.847(dd,1H,J=3.2,9.0 Hz), 7.325-7.356(m,2H), 7.435(d,1H,J=7.8 Hz),7.852(s,1H), 7.915(d,1H,J=8.8 Hz), 9,202(s,1H)

Fab-MS(m/z);537(M+1)⁺

Example 9

Compound 11

To a solution of 5 mg (0.01 mmol) of 2-methyl-17-formylstaurosporine in0.2 ml of dichloroethane was added 7 mg of(carboethoxymethylene)triphenylphosphorane and the mixture was stirredat room temperature overnight, followed by stirring at 50° C. for 6hours. The reaction mixture was washed with brine and dried over K₂ CO₃.The solvent was removed by evaporation and the residue was purified bypreparative TLC (MeOH/CHCl₃ 1/9) to give 5 mg (86%) of Compound 11.

¹ HNMR(DMSO-d₆) δ; 1.312(t,3H,J=7.1 Hz), 1.420(br.s,3H), 2.315(s,3H),3.394(s,3H), 4.080(d,1H,J=3.5 Hz), 4.229(q,2H,J=7.1 Hz), 5.058(s,2H),6,571(d,1H,J=15.8 Hz), 6.751(m,1H), 7.281-8.002(m,7H), 9.581(d,1H,J=1.5Hz).

Fab-MS(m/z); 579(M+1)⁺

Example 10

Compound 12

To 20 ml of CH₂ Cl₂ cooled to 0° C., 108 μl (1.22 ml) oftrifluoromethanesulfonic acid and 52 μl (1.22 mmol) of fuming nitricacid were added, followed by stirring for 20 minutes. To the reactionmixture cooled to -78° C. was added dropwise, a solution of 500 mg (0.81mmol) of 11-Cbz-staurosporine in 5 ml of CH₂ Cl₂, followed by stirringfor 1 hour. The reaction was stopped by the addition of a saturatedaqueous solution of sodium hydrogen carbonate. The reaction mixture wasextracted with CHCl₃ and the extract was washed with water and brine,and then dried over Na₂ SO₄. After the solvent was removed byevaporation, the residue was purified by silica gel columnchromatography (MeOH/CHCl₃ 1/100) to give 537 mg (99%) of 17-nitroderivative.

¹ HNMR(DMSO-d₆) δ; 2.125-2.232(m,1H), 2.322(s,3H), 2.635(s,3H),2.708(s,3H), 3.300(s,3H), 3.394(br.s,1H), 4.602(br.s,1H), 5.020(s,2H),5.186(s,2H), 7.104(br.s,1H), 7.362-7.450(m,6H), 7.543(t,1H,J=7.9 Hz),7.834(d,1H,J=9.1 Hz), 8.123(d,1H,J=8.0 Hz), 8.362(dd,1H,J=2.2,9.0 Hz),8.724(s,1H), 10.231(d,1H,J=2.4 Hz)

Fab-MS(m/z);660(M+1)⁺

Compound 12 was obtained from 200 mg (0.30 mmol) of the 17-nitroderivative and 100 mg of 10% Pd/C by the procedure analogous to that ofExample 8 (yield: 145 mg, 84%).

¹ HNMR(DMSO-d₆) δ; 2.066-2.183(m,1H), 2.283(s,3H), 2.675(s,3H),3.284(s,3H), 4.043(br.s,1H), 4.562(s,1H), 5.112(s,2H),6.990(dd,1H,J=2.8,9.5 Hz), 7.429(t,1H,J=7.5 Hz), 7.517-7.587(m,2H),7.704(d,1H,J=10.7 Hz), 8.088-8.174(m,2H), 9.163(br.s,1H),9.407(d,1H,J=2.1 Hz), 10.204(br.s,1H)

Fab-MS(m/z);496(M+1)⁺

Example 11

Compound 13

A solution of 518 mg (0.80 mmol) of2-methyl-11-Cbz-17-nitrostaurosporine in 25 ml of THF was cooled to 0°C. and 74 mg (1.1 mmol) of zinc powder and 2.5 ml of 2N HCl were addedthereto, followed by stirring for 30 minutes. The mixture was furtherstirred at room temperature for 1 day. After the completion of reaction,the reaction mixture was filtered. A saturated aqueous solution ofsodium hydrogen carbonate was added to the filtrate, followed byextraction with CHCl₃. The extract was washed with water and brine, andthen dried over Na₂ SO₄. After the solvent was removed by evaporation,the residue was purified by silica gel column chromatography (MeOH/CHCl₃1/50) to give 271 mg (54%) of 11-Cbz derivative of Compound 12.

¹ HNMR(DMSO-d₆) δ; 2.333(br.s,3H), 2.671(br.s,6H), 2.737(s,3H),3.257(s,3H), 4.254(br.s,1H), 4.653(br.s,1H), 5.029(s,2H), 5.187(s,2H),6.849(br.s,1H), 6.893(dd,1H,J=2.2,8.6 Hz), 7.324-7.482(m,8H),8.019(d,1H,J=7.7 Hz), 8.554(s,1H)

Fab-MS(m/z);630(M+1)⁺

To a solution of 200 mg (0.32 mmol) of the 11-Cbz derivative of Compound12 in 10 ml of THF and 1 ml of water, was added 200 mg (3.2 mmol) ofsodium cyanoborohydride and 890 μl (31.8 mmol) of formaldehyde. Themixture was adjusted to pH 5-6 with 2N hydrochloric acid, and stirred atroom temperature for 1.5 hours. After the completion of reaction, thereaction mixture was diluted with CHCl₃ and washed successively with asaturated aqueous solution of NaHCO₃, water and brine, and then driedover Na₂ SO₄. The solvent was removed by evaporation and the residue waspurified by silica gel column chromatography (MeOH/CHCl₃ 1/50) to give88 mg (42%) of 11-Cbz derivative of Compound 13.

¹ HNMR(CDCl₃) δ; 1.750(br.s,3H), 2.184(br.s,1H), 2.469(s,3H),2.496(s,3H), 2.802(s,3H), 3.137(s,3H), 3.389(s,3H), 4.036(br.s,1H),4.651(br.s,1H), 4.947(s,2H), 5.190(s,2H), 6.678(br.s,1H),7.184-7.516(m,8H), 7.735(br, d,1H,J=6.9 Hz), 7.927(s,1H), 7.945(s,1H),9.221(br.s,1H)

Fab-MS(m/z);658(M+1)⁺

Compound 13 was obtained from 102 mg (0.16 mmol) of the 11-Cbzderivative of Compound 13 and 50 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 73 mg, 78%).

¹ HNMR(DMSO-d₆) δ; 2.258(s,3H), 2.667(s,6H), 2.735(s,3H), 2.893(s,3H),3.284(s,3H), 4.038(br.s,1H), 4.565(s,1H), 5.106(d,1H,J=18.0 Hz),5.151(d,1H,J=18.2 Hz), 6.983(dd,1H,J=3.2,9.4 Hz), 7.447(t,1H,J=7.4 Hz),7.588(t,1H,J=7.4 Hz), 7.768(br.s,1H), 7.887(br.s,1H), 7.953(s,1H),8.094(d,1H,J=3.1 Hz), 8.106(s,1H), 9.201(br.s,1H), 9.522(br.s,1H)

Fab-MS(m/z);524(M+1)⁺

Example 12

Compound 14

A solution of 87 mg (0.41 mmol) of N-Cbz-glycine in 10 ml of CH₂ Cl₂ wascooled to 0° C. and 87 mg (0.41 mmol) of3-(3-diethylaminopropyl)-1-ethylcarbodiimide.hydrochloride and 200 mg(0.32 mmol) of the 11-Cbz derivative of Compound 12 were added thereto,followd by stirring under an argon atmosphere at room temperature for 1hour. The reaction was stopped by an addition of ice water, and thereaction mixture was extracted with CHCl₃. The extract was washed with asaturated aqueous solution of NaHCO₃ and brine, and then dried over Na₂SO₄. The solvent was removed by evaporation,and the residue was purifiedby silica gel column chromatography (MeOH/CHCl₃ 3/100) to give 213 mg(82%) of 17-(N-Cbz-glycyl)amino derivative.

¹ HNMR(DMSO-d₆, 90° C.) δ; 2.302(s,3H), 2.625(s,3H), 2.749(s,3H),3.276(s,3H), 3.927(d,2H,J=6.1 Hz), 4.649-4.699(m,1H), 5.031(s,2H),5.101(s,2H), 5.194(d,1H,J=12.4 Hz), 5.253(d,1H,J=12.5 Hz),6.927(dd,1H,J=5.9,8.8 Hz), 7.094(br.s,1H), 7.262-7.517(m,13H),7.827(d,1H,J=8.8 Hz), 7.893(d,1H,J=8.3 Hz), 8.041(d,1H,J=7.8 Hz),9.303(d,1H,J=2.2 Hz), 9.748(s,1H)

Fab-MS(m/z);821(M+1)⁺

Compound 14 was obtained from 213 mg (0.26 mmol) of the17-(N-Cbz-glycyl)amino derivative and 106 mg of 10% Pd/C by theprocedure analogous to that of Example 8 (yield: 113 mg, 70%).

¹ HNMR(DMSO-d₆) δ;2.067-2.136(m,1H), 2.319(s,3H), 2.672(s,3H),3.273(s,3H), 3.847(d,2H,J=5.6 Hz), 4.038(br.s,1H), 4.581(s,1H),5.091(s,2H), 6.942(dd,1H,J=3.2,9.5 Hz), 7.433(t,1H,J=7.6 Hz),7.535-7.571(m,2H), 7.896(dd,1H,J=2.2,8.8 Hz), 8.070-8.097(m,2H),8.235(br.s,2H), 9.436(d,1H,J=1.9 Hz), 9.199(br.s,1H), 9.523(br.s,1H),10.622(s,1H)

Fab-MS(m/z);553(M+1)⁺

Example 13

Compound 15

A solution of 250 mg (0.40 mmol) of the 11-Cbz derivative of Compound 12in 10 ml of CH₂ Cl₂ was cooled to 0° C. and 55 μl (0.40 mmol) oftriethylamine and 35 μl (0.44 mmol) of chloroacetyl chloride were addedthereto, followed by stirring for 30 minutes. After the completion ofreaction, the reaction mixture was diluted with CHCl₃, washed with waterand brine, and then dried over Na₂ SO₄. The solvent was removed byevaporation, and the residue was purified by silica gel columnchromatography (MeOH/CHCl₃ 1/100) to give 190 mg (69%) of17-chloroacetylamino derivative.

¹ HNMR(DMSO-d₆) δ; 2.201-2.281(m,1H), 2.332(br.s,3H), 2.670(s,3H),2.743(s,3H), 3.276(s,3H), 4.274(br.s,1H), 4.309(s,1H), 4.672(br.s,1H),5.077(s,2H), 5.195(s,2H), 6.972(br.s,1H), 7.312-7.387(m,7H),7,503(t,1H,J=7.8 Hz), 7.596(d,1H,J=8.8 Hz), 7.894(dd,1H,J=2.2,8.8 Hz),8.298(d,1H,J=7.3 Hz), 9.303(d,1H,J=2.2Hz), 10.418(s,1H)

Fab-MS(m/z);706(M+1)⁺

To a solution of 150 mg (0.21 mmol) of the 17-chloroacetyl-aminoderivative in 10 ml of CHCl₃ were added successively 148 μl (1.1 mmol)of triethylamine, 185.2 μl (1.1 mmol) of diisopropylethylamine and 118μl (1.1 mmol) of N-methylpiperazine, followed by stirring at 60° C. for9 hours. After the completion of reaction, the reaction mixture wasdiluted with CHCl₃, washed with water and brine, and then dried over Na₂SO₄. The solvent was removed by evaporation, and the residue waspurified by silica gel column chromatography (MeOH/CHCl₃ 7.5/100) togive 128 mg (78%) of 11-Cbz derivative of Compound 15.

¹ HNMR(DMSO-d₆) δ; 2.215(s,3H), 2.738(s,3H), 3.171(s,2H), 3.273(s,3H),4.279(br.s,1H), 4.681(br.s,1H), 5.074(s,2H), 5.194(s,2H),6.970(br.s,1H), 7.352-7.399(m,6H), 7.486-7.515(m,1H), 7. 569(d,1H,J=8.5Hz), 7.832(dd,1H,J=2.1,8.8 Hz), 8.054(d,1H,J=7.7 Hz), 9.268(d,1H,J=2.0Hz), 9.720(s,1H)

Fab-MS(m/z);770(M+1)⁺

Compound 15 was obtained from 128 mg (1.17 mmol) of the 11-Cbzderivative of Compound 15 and 64 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 103 mg, 83%).

¹ HNMR(DMSO-d₆) δ; 2.070-2.131(m,1H), 2.313(s,3H), 2.672(s,3H),2.856(s,3H), 3.275(s,3H), 4.020-4.032(m,1H), 4.586(s,1H), 5.093(s,2H),6.942(dd,1H,J=3.3,9.4 Hz), 7.414(t,1H,J=7.6 Hz), 7.553(s,1H),7.555(s,1H), 7.870(dd,1H,J=2.2,8.7 Hz), 8.080(d,1H,J=3.9 Hz),8.098(d,1H,J=2.4 Hz), 9.220(br.s,1H), 9.431(s,1H), 9.549(br.s,1H),10.451(br.s,1H), 10.247(br.s,1H)

Fab-MS(m/z);636(M+1)⁺

Example 14

Compound 16

5,17-Dinitro derivative was obtained from 500 mg (0.81 mmol) of2-methyl-11-Cbz-staurosporine, 540 μl (6.11 mmol) oftrifluoromethanesulfonic acid and 258 μl (6.11 mmol) of fuming nitricacid by the procedure analogous to that of Example 10 (yield: 200 mg,35%).

¹ HNMR(DMSO-d₆) δ; 2.115-2.200(m,1H), 2.361(s,3H), 2.617(s,3H),2.711(s,3H), 3.255(s,3H), 3.372(br.s,1H), 4.635(br.s,1H), 5.188(s,4H),7.087(t,1H,J=7.1 Hz), 7.409(br.s,5H), 7.776(d,1H,J=9.0 Hz),8.189(br.s,1H), 8.287(dd,1H,J=2.4,9.1 Hz), 8.366(d,1H,J=8.8 Hz),8.781(d,1H,J=2.4 Hz), 10.148(d,1H,J=2.4 Hz)

Fab-MS(m/z);705(M+1)⁺

Compound 16 was obtained from 175 mg (0.25 mmol) of the 5,17-dinitroderivative and 88 mg of 10% Pd/C by the procedure analogous to that ofExample 8 (yield: 135 mg, 94%).

¹ HNMR(DMSO-d₆) δ; 2.054-2.149(m,1H), 2.295(s,3H), 2.671(s,3H),4.036(s,1H), 4.556(s,1H), 5.061(s,2H), 6.986(dd,1H,J=3.2,9.0 Hz),7.455(d,1H,J=7.1 Hz), 7.542(dd,1H,J=2.2,8.7 Hz), 7.708(d,1H,J=8.7 Hz),7.969(s,1H), 8.153(d,1H,J=9.1 Hz), 9.395(d,1H,J=2.0 Hz).

Fab-MS(m/z);511(M+1)⁺

Example 15

Compound 17

11-Cbz derivative of Compound 16 was obtained from 1.0 g of2-methyl-11-Cbz-5,17-dinitrostaurosporine, 0.93 g of zinc powder and 10ml of 2N HCl by the procedure analogous to that of Example 11 (yield:573 mg, 63%).

¹ HNMR(DMSO-d₆) δ;2.216-2.205(m,1H), 2.263(s,3H), 2.624(s,3H),2.727(s,3H), 3.246(s,3H), 4.126(br.s,1H), 4.929(s,2H), 5.001(br.s,2H),5.146(br.s,2H), 5.176(s,2H), 6.800-6.822(m,1H), 6.843(d,1H,J=2.2 Hz),6.864(d,1H,J=2.2 Hz), 7.181(d,1H,J=2.1 Hz), 7.286(d,1H,J=8.8 Hz),7.396(br.s,5H), 7.679-7.699(m,1H), 8.530(d,1H,J=2.0 Hz)

Fab-MS(m/z);645(M+1)⁺

11-Cbz derivative of Compound 17 was obtained from 200 mg (0.31 mmol) ofthe 11-Cbz derivative of Compound 16, 195 mg (3.10 mmol) of sodiumcyanoborohydride and 861 μl (31.0 mmol) of formaldehyde by the procedureanalogous to that of Example 11 (yield: 97 mg, 45%).

¹ HNMR(DMSO-d₆) δ; 2.626(s,3H), 2.727(s,3H), 3.045(s,6H), 3.068(s,3H),3.277(s,3H), 4.206(br.s,1H), 4.658(br.s,1H), 5.079(s,2H), 5.185(s,2H),6.922(br.s,1H), 7.135(br.s,1H), 7.332(br.s,5H), 7.547(br.s,2H),7.665-7.722(m,1H), 7.841(br.s,1H), 8.076(s,1H)

Fab-MS(m/z);701(M+1)⁺

Compound 17 was obtained from 97 mg (0.14 mmol) of the 11-Cbz derivativeof Compound 17 and 100 mg of 10% Pd/C by the procedure analogous to thatof Example 8 (yield: 60 mg, 68%).

¹ HNMR(DMSO-d₆,90° C.) δ; 2.113-2.167(m,1H), 2.383(s,3H), 2.658(s,3H),3.164(s,3H), 3.220(s,3H), 3.304(s,2H), 3.904(br.s,1H), 4.578(s,1H),5.105(s,2H), 6.922-6.937(m,1H), 7.517(br.s,1H), 7.681(br.d,1H,J=9.0 Hz),7.747(br.s,1H), 7.855(br.s,1H), 8.031(d,1H,J=9.3 Hz), 9.135(br.s,2H),9.494(s,1H), 9.641(br.s,1H)

Fab-MS(m/z);567(M+1)⁺

Example 16

Compound 18

5,17-Bis(N-Cbz-glycylamino) derivative was obtained from 200 mg (0.31mmol) of the 11-Cbz derivative of Compound 16, 149 mg (0.71 mmol) ofN-Cbz-glycine and 86 mg (0.71 mmol) of3-(3-diethylaminopropyl)-1-ethylcarbodiimide.hydrochloride by theprocedure analogous to that of Example 12 (yield: 198 mg, 62%).

¹ HNMR(DMSO-d₆,90° C.) δ; 2.267-2.280(m,1H), 2.293(s,3H), 2.633(s,3H),2.744(s,3H), 3.284(s,3H), 3.908(d,2H,J=2.7 Hz), 3.924(d,2H,J=2.7 Hz),4.181(s,1H), 4.641-4.680(m,1H), 4.985(s,2H), 5.097(s,2H), 5.101(s,2H),5.195(d,1H,J=12.5 Hz), 5.246(d,1H,J=12.5 Hz), 6.931(dd,1H,J=5.8,8.7 Hz),7.169(br.s,2H), 7.268-7.450(m,15H), 7.521(d,1H,J=8.8 Hz),7.661(dd,1H,J=2.2,9.0 Hz), 7.817(dd,1H,J=2.0,8.8 Hz), 7.851(d,1H,J=9.3Hz), 8.349(d,1H,J=2.0 Hz), 9.289(d,1H,J=2.0 Hz), 9.748(s,1H),9.824(s,1H)

Fab-MS(m/z);1027(M+1)⁺

Compound 18 was obtained from 198 mg (0.19 mmol) of the5,17-bis(N-Cbz-glycylamino) derivative and 100 mg of 10% Pd/C by theprocedure analogous to that of Example 8 (yield: 81 mg, 57%).

¹ HNMR(DMSO-d₆, 90° C.) δ; 2.114-2.307(m,1H), 2.422(s,3H), 2.644(s,3H),3.279(s,3H), 3.821(s,2H), 3.856(s,2H), 3.921(br.s,1H), 4.591(s,1H),4.994(s,2H), 6.903(dd,1H,J=3.4,9.3 Hz), 7.532(d,1H,J=8.6 Hz),7.727(dd,1H,J=2.2,9.0 Hz), 7.831(d,1H,J=8.3 Hz), 8.013(d,1H,J=9.0 Hz),8.206(br.s,3H), 8.444(d,1H,J=2.0 Hz), 9.426(d,1H,J=1.7 Hz),10.422(s,1H), 10.756(s,1H)

Fab-MS(m/z); 625(M+1)⁺

Example 17

Compound 19

To a solution of 200 mg (0.31 mmol) of the 11-Cbz derivative of Compound16 in 10 ml of CH₂ Cl₂ was added 91.6 μl (1.6 mmol) of methyl isocyanateand the mixture was heated under reflux for 2 hours. After thecompletion of reaction, the reaction mixture was diluted with CHCl₃,washed with water and brine, and then dried over Na₂ SO₄. The solventwas removed by evaporation, and the residue was purified by silica gelcolumn chromatography (MeOH/CHCl₃ 1/20) to give 147 mg (63%) of 11-Cbzderivative of Compound 19.

¹ HNMR(DMSO-d₆,90° C.) δ; 2.266(s,3H), 2.622(s,3H), 2.699(s,3H),2.723(s,3H), 2.741(s,3H), 3.274(s,3H), 4.144(s,1H), 4.640-4.668(m,1H),4.693(s,2H), 5.188(d,1H,J=12.5 Hz), 5.243(d,1H,J=12.7 Hz),5.783(br.s,1H), 5.862(br.s,1H), 6.882(dd,1H,J=5.8,8.8 Hz),7.325-7.442(m,7H), 7.751(d,1H,J=9.8 Hz), 7.778(dd,1H,J=2.2,8.8 Hz),8.138(d,1H,J=2.2 Hz), 8.230(br.s,1H), 8.321(br.s,1H), 9.029(d,1H,J=2.2Hz)

Fab-MS(m/z); 759(M+1)⁺

Compound 19 was obtained from 147 mg (0.19 mmol) of the 11-Cbzderivative of Compound 19 and 150 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 104 mg, 81%).

¹ HNMR(DMSO-d₆, 90° C.) δ; 2.086-2.157(m,1H), 2.393(s,3H), 2.437(s,3H),2.666(s,3H), 2.702(s,3H), 2.724(s,3H), 3.274(s,3H), 3.936(br.s,1H),4.452(s,1H), 4.971(s,2H), 6.842(dd,1H,J=3.4,9.3 Hz), 7.397(d,1H,J=8.8Hz), 7.482(dd,1H,J=2.2,9.0 Hz), 7.837(dd,1H,J=2.2,9.0 Hz),7.866(d,1H,J=9.0 Hz), 8.191(d,1H,J=2.2 Hz), 8.332(br.s,1H),8.494(br.s,1H), 8.818(br.s,1H), 9.051(d,1H,J=2.0 Hz), 9.198(br.s,1H)

Fab-MS(m/z);625(M+1)⁺

Example 18

Compound 20

5,17-Bis(chloroacetylamino) derivative was obtained from 200 mg (0.31mmol) of the 11-Cbz derivative of Compound 16, 87 μl (0.62 mmol) oftriethylamine and 57 μl (0.71 mmol) of chloroacetyl chloride by theprocedure analogous to that of Example 13 (yield: 129 mg, 52%).

¹ HNMR(DMSO-d₆) δ; 2.193-2.365(m,1H), 2.661(s,3H), 2.738(s,3H),4.231(br.s,1H), 4.308(s,2H), 4.323(s,2H), 4.671(br.s,1H), 5.031(s,2H)5.195(s,2H), 6.974(br.s,1H), 7.399(br.s,5H), 7.586(d,1H,J=8.5 Hz), 7.684(d,1H,J=9.1 Hz), 7 893(dd,1H,J=2.0,8.8 Hz), 8.383(d,1H,J=2.0 Hz),9.296(d,1H,J=2.0 Hz), 10.413(s,1H), 10.437(s,1H)

Fab-MS(m/z);797(M)⁺

11-Cbz derivative of Compound 20 was obtained from 129 mg (0.16 mmol) ofthe 5,17-bis(chloroacetylamino) derivative, 226 μl (1.6 mmol) oftriethylamine, 283 μl (1.6 mmol) of diisopropylethylamine and 180 μl(1.6 mmol) of N-methylpiperazine by the procedure analogous to that ofExample 13 (yield: 138 mg, 92%).

¹ HNMR(DMSO-d₆,90° C.) δ; 2.222(s,6H), 2.287(s,3H), 2.434-2.456(m,4H),2.554-2.613(m,4H), 2.744(s,3H), 3.163(s,2H), 3.180(s,2H), 3.289(s,3H),4.184(s,1H), 4.682(br.d,J=11.5 Hz), 5.000(s,2H), 5.193(d,1H,J=12.5 Hz),5.246(d,1H,J=12.5 Hz), 6.933(dd,1H,J=5.8,8.6 Hz), 7.345-7.449(m,5H),7.525(d,1H,J=8.8 Hz), 7.680(dd,1H,J=2.0,9,0 Hz), 7.783-7.839(m,2H),8.362(d,1H,J=1.7 Hz), 9.264(d,1H,J=1.7 Hz), 9.466(s,1H), 9.584(s,1H)

Fab-MS(m/z);925(M+1)⁺

Compound 20 was obtained from 128 mg (0.14 mmol) of the 11-Cbzderivative of Compound 20, and 130 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 113 mg, 84%).

¹ HNMR(DMSO-d₆,90° C.) δ; 2.417(s,3H), 2.467(s,3H), 2.663(s,3H),2.777(s,6H), 3.289(s,3H), 3.511(s,2H), 3.552(s,2H), 3.942(br.s,1H),4.578(s,1H), 5.009(s,2H), 6.898(dd,1H,J=3.7,9.3 Hz), 7.504(d,1H,J=8.8Hz), 7.747(dd,1H,J=2.2,9,0 Hz), 7.827(dd,1H,J=2.0,8.8 Hz),7.977(d,1H,J=9.0 Hz), 8.442(d,1H,J=2.2 Hz), 9.071(br.s,1H),9.360(d,1H,J=2.0 Hz), 9.633(br.s,1H), 9.902(s,1H)

Fab-MS(m/z);791(M+1)⁺

Example 19

Compound 21

To a solution of 4.5 g (6.44 mmol) of 2-methyl-11-Cbz-5,17-diacetylstaurosporine in 450 ml of CH₂ Cl₂ was added 2.17 g (25.8 mmol) ofsodium hydrogen carbonate and 8.34 g (48.4 mmol) of 55%m-chloroperbenzoic acid, followed by stirring at room temperature for 5hours. After the completion of reaction, the reaction mixture wasdiluted with CHCl₃, washed succesively with a saturated aqueous solutionof sodium sulfite, a saturated aqueous solution of sodium hydrogencarbonate and water, and then dried over Na₂ SO₄. The solvent wasremoved by evaporation and the residue was dissolved in 250 ml of CH₂Cl₂ and cooled to 0° C. 3.15 ml (12.9 mmol) of 28% sodium methoxide inMeOH was added thereto followed by stirring for 10 minutes. After thecompletion of reaction, the reaction mixture was neutralized with 2NHCl, followed by extraction with CHCl₃. The extract was washed withbrine, and then dried over Na₂ SO₄. The solvent was removed byevaporation, and the residue was purified by silica gel columnchromatography (MeOH/CHCl₃ 1/20) to give 2.67 g (63%) of 5,17-dihydroxyderivative.

¹ HNMR(DMSO-d₆) δ; 2.635(s,3H), 2.733(s,3H), 4.137(br.s,1H),4.635(br.s,1H), 4.977(s,2H), 5.181(s,2H), 6.855(br.s,1H),6.937-6.973(m,2H), 7.300-7.571(m,6H), 7.646-7.718(m,2H), 8.716(s,1H),9.007(s,1H), 9.208(s,1H)

Fab-MS(m/z);646(M)⁺

11-Cbz derivative of Compound 21 was obtained from 50 mg (0.077 mmol) ofthe 5,17-dihydroxy derivative, 54 μl (0.39 mmol) of triethylamine and27.5 μl (0.39 mmol) of acetyl chloride by the procedure analogous tothat of Example 13 (yield: 52 mg, 92%).

¹ NMR(DMSO-d₆) δ; 2.213-2.327(m,1H), 2.353(s,6H), 2.690(s,3H),2.745(s,3H), 3.244(s,3H), 4.298(br.s,1H), 4.663(br.s,1H), 5.047(s,2H),5.198(s,2H), 7.006(br.s,1H), 7.251(d,1H,J=2.4 Hz), 7.273(d,1H,J=2.4 Hz),7.400(br.s,5H), 7.623(d,1H,J=8.8 Hz), 7.801(d,1H,J=2.5 Hz),7.994(br.s,1H), 8.994(d,1H,J=2.4 Hz)

Fab-MS(m/z);731(M+1)⁺

Compound 21 was obtained from 150 mg (0.21 mmol) of the 11-Cbzderivative of Compound 21 and 75 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 120 mg, 92%).

¹ HNMR(DMSO-d₆) δ; 2.045-2.134(m,1H), 2.273(s,3H), 2.363(s,6H),2.669(s,3H), 3.239(s,3H), 4.022(br.s,1H), 4.510(s,1H), 5.003(s,2H),6.925(dd,1H,J=3.1,9.4 Hz), 7.274-7.386(m,2H), 7.580(d,1H,J=8.8 Hz),7.803(d,1H,J=2.4 Hz), 8.106(d,1H,J=9.0 Hz), 9.024(d,1H,J=2.5 Hz)

Fab-MS(m/z);597(M+1)⁺

Example 20

Compound 22

11-Cbz derivative of Compound 22 was obtained from 150 mg (0.23 mmol) of2-methyl-11-Cbz-5,17-dihydroxy staurosporine, 162 μl (1.2 mmol) oftriethylamine and 143 μl (1.2 mmol) of pivaloyl chloride by theprocedure analogous to that of Example 19 (yield: 113 mg, 60%).

¹ HNMR(DMSO-d₆, 90° C.) δ; 1.391(s,9H), 1.402(s,9H), 2.302(s,3H),2.673(s,3H), 2.755(s,3H), 3.259(s,3H), 4,231(s,1H), 4.650-4.700(m,1H),5.033(s,2H), 5.197(d,1H,J=12.5 Hz), 5.248(d,1H,J=12.2 Hz),6.981(dd,1H,J=6.0,8.7 Hz), 7.168-7.224(m,2H), 7.325-7 450(m,7H),7.615(d,1H,J=8.5 Hz), 7.719(d,1H,J=2.2 Hz), 7.929(d,1H,J=9.3 Hz),9.005(d,1H,J=2.4 Hz)

Fab-MS(m/z);815(M+1)⁺

Compound 22 was obtained from 110 mg (0.13 mmol) of the 11-Cbzderivative of Compound 22 and 50 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 85 mg, 88%).

¹ HNMR(DMSO-d₆, 90° C.) δ; 1.396-1.404(m,18H), 2.187-2.246(m,1H),3.260(s,3H), 3.888(br.s,1H), 4.468(s,1H), 5.035(s,2H),6.904(dd,1H,J=3.3,8.9 Hz), 7.193-7.252(m,2H), 7.571(d,1H,J=8.6 Hz),7.736(d,1H,J=2.4 Hz), 8.045(d,1H,J=9.0 Hz), 9.030(d,1H,J=2.4 Hz)

Fab-MS(m/z);681(M+1)⁺

Example 21

Compound 23

11-Cbz derivative of Compound 23 was obtained from 150 mg (0.23 mmol) of2-methyl-11-Cbz-5,17-dihydroxystaurosporine, 162 μl (1.2 mmol) oftriethylamine and 319 μ(1.2 mmol) of palmitoyl chloride by the procedureanalogous to that of Example 19 (yield: 235 mg, 90%).

¹ HNMR(DMSO-d₆, 90° C.) δ;0.874(t,6H,J=6.8,6.8 Hz), 1.259-1.481(m,48H),1.717-1.780(m,4H), 2.316(s,3H), 2.618-2.655(m,4H), 2.667(s,3H),2.766(s,3H), 3.261(s,3H), 4.233(s,1H), 4.675-4.702(m,1H), 5.013(s,2H),5.210(d,1H,J=12.5 Hz), 5.261(s,1H,J=12.5 Hz), 6.973(dd,1H,J=5.9,8.5 Hz),7.229(dd,1H,J=2.4,8.8 Hz), 7.260(dd,1H,J=2.4,9.2 Hz), 7.356-7.463(m,5H),7.607(d,1H,J=8.5 Hz), 7.751(d,1H,J=2.2 Hz), 7.931(d,1H,J=9.3 Hz),9.041(d,1H,J=2.6 Hz)

Fab-MS(m/z); 1123(M+1)⁺

Compound 23 was obtained from 235 mg (0.21 mmol) of the 11-Cbzderivative of Compound 23 and 120 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 193 mg, 90%).

¹ HNMR(DMSO-d₆, 90° C.) δ;0.846(t,1H,J=6.5 Hz), 1,248-1.703(m,48H),1.703-1.784(m,4H), 2.156-2.225(m,1H), 2.612-2.657(m,4H), 3.251(s,3H),3.948(br.s,1H), 4.474(s,1H), 5.014(s,2H), 6.915(dd,1H,J=3.4,9.0 Hz),7.251(dd,1H,J=2.2,8.8 Hz), 7.287(dd,1H,J=2.4,9.3 Hz), 7.562(d,1H,J=8.3Hz), 7.771(d,1H,J=2.4 Hz), 8.045(d,1H,J=9.0 Hz), 9.048(d,1H,J=2.0 Hz)

Fab=MS (m/z);989(M+1)⁺

Example 22

Compound 24

A solution of 150 mg (0.23 mmol) of2-methyl-11-Cbz-5,17-dihydroxystaurosporine in 10 ml of THF was cooledto 0° C. and 101 μl (1.2 mmol) of chlorosulfonyl isocyanate was addedthereto, followed by stirring for 1.5 hours. To the reaction mixturewere added 10 ml of THF and 3 ml of water and stirred at 70° C. for 1hour. After the completion of reaction, the reaction mixture was dilutedwith CHCl₃ and washed with brine, and then dried over Na₂ SO₄. Thesolvent was removed by evaporation, and the residue was purified bysilica gel column chromatography (MeOH/CHCl₃ 3/20) to give 110 mg (65%)of 11-Cbz derivative of Compound 24.

¹ HNMR(DMSO-d₆) δ; 2.344(br.s,3H), 2.689(s,3H), 2.744(s,3H),3.251(s,3H), 4.272(br.s,1H), 4.680(br.s,1H), 5.061(s,2H), 5.195(s,2H),6.992(br.s,1H), 7.220(d,1H,J=2.4 Hz), 7.237(d,1H,J=2.4 Hz),7.398(br.s,5H), 7.598(d,1H,J=8.8 Hz), 7.710(d,1H,J=2.4 Hz),7.960(d,1H,J=8.8 Hz), 8.971(d,1H,J=2.4 Hz)

Fab-MS(m/z);733(M+1)⁺

Compound 24 was obtained from 110 mg (0.15 mmol) of the 11-Cbzderivative of Compound 24 and 100 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 76 mg, 80%).

¹ HNMR(DMOS-d₆) δ;2.075-2.143(m,1H), 2.294(s,3H), 2.671(s,3H),3.249(s,3H), 4.022(br.s,1H), 4.501(s,1H), 5.051(s,2H),6.931(dd,1H,J=2.9,9.3 Hz), 7.240-7.293(m,2H), 7.540(d,1H,J=8.5 Hz),7.742(d,1H,J=2.5 Hz), 8.052(d,1H,J=9.0 Hz), 9.000(d,1H,J=2.4 Hz),9.232(br.s,1H)

Fab-MS(m/z);599(M+1)⁺

Example 23

Compound 25

A solution of 300 mg (0.46 mmol) of2-methyl-11-Cbz-5,17-dihydroxystaurosporine in 15 ml of THF was cooledto 0° C. and 324 μl (2.3 mmol) of triethyl amine and 468 mg (2.3 mmol)of 4-nitrophenyl chloroformate were added thereto, followed by stirringfor 8 hours. After the completion of reaction, the reaction mixture wasdiluted with CHCl₃ and washed with a saturated aqueous solution ofsodium hydrogen carbonate and brine then dried over Na₂ SO₄. The solventwas removed by evaporation, and the residue was purified by preparativeTLC (MeOH/CHCl₃ 1/100) to give 6 mg (26%) of5,17-bis(4-nitrophenyloxycarbonyl) derivative.

¹ HNMR(DMSO-d₆) δ; 2.720(s,3H), 2.751(s,3H), 3.275(s,3H),4.311(br.s,1H), 4.672(br.s,1H), 5.100(s,2H), 5.204(s,2H),7.055(t,1H,J=7.2Hz), 7.410(br.s,5H), 7.582(d,1H,J=2.5 Hz),7.604(d,1H,J=2.4 Hz), 7.733(s,1H), 7.749-7.795(m,4H), 8.122(d,1H,J=2.7Hz), 8.362-8.420(m,4H), 9.236(d,1H,J=2.5 Hz)

Fab-MS(m/z);977(M+1)⁺

To a solution of 150 mg (0.15 mmol) of the5,17-bis(4-nitrophenyloxycarbonyl) derivative in 10 ml of CHCl₃ wereadded 107 μl (0.77 mmol) of triethylamine and 85 μl (0.77 mmol) ofN-methylpiperazine and the mixture was stirred at room temperature for 4hours. After the completion of reaction, the reaction mixture wasdiluted with CHCl₃ and washed successively with a saturated aqueoussolution of sodium hydrogen carbonate, water and brine, and dried overNa₂ SO₄. The solvent was removed by evaporation, and the residue waspurified by silica gel column chromatography (MeOH/CHCl₃ 1/10) to give121 mg (88%) of 11-Cbz derivative of Compound 25.

¹ HNMR(DMSO-d₆) δ; 2.265(s,9H), 2.427(br.s,8H), 2.675(s,3H),2.742(s,4H), 3.245(s,3H), 3.487(br.s,4H), 3.683(br.s,4H),4.264(br.s,1H), 4.672(br.s,1H), 5.050(s,2H), 5.194(s,2H),6.985(br.s,1H), 7.232(d,1H,J=2.5 Hz), 7.254(d,1H,J=2.4 Hz),7.402(br.s,5H), 7.610(d,1H,J=8.8 Hz), 7.748(d,1H,J=2.4 Hz),7.957(br.s,1H), 8.976(br.s,1H)

Fab-MS(m/z);899(M+1)⁺

Compound 25 was obtained from 121 mg (0.13 mol) of the 11-Cbz derivativeof Compound 25 and 60 mg of 10% Pd/C by the procedure analogous to thatof Example 8 (yield: 94 mg, 80%).

¹ HNMR(DMSO-d₆) δ; 2.411(s,3H), 2.664(s,3H), 2.833(s,3H), 2.837(s,3H),3.259(s,3H), 3.467(br.s,4H), 3.954(br.s,1H), 4.234(br.s,4H),4.608(s,1H), 5.038(s,2H), 6.937(dd,1H,J=3.4,9.3 Hz),7.315(dd,1H,J=2.5,8.8 Hz), 7.367(dd,1H,J=2.4,9.0 Hz), 7.571(d,1H,J=8.8Hz), 7.838(d,1H,J=2.4 Hz), 8.056(d,1H,J=9.3 Hz), 9.055(br.s,1H),9.090(d,1H,J=2.2 Hz), 9.626(br.s,1H), 11.342(br.s,1H)

Fab-MS(m/z);765(M+1)⁺

Example 24

Compound 26

To a solution of 5 mg (0.009 mmol) of Compound 11 in 0.2 ml of DMF wereadded 3 mg of 20% Pd(OH)₂ /C and 0.1 ml of acetic acid and the mixturewas stirred under hydrogen atmosphere at room temperature for 1 hour.The reaction mixture was filtered with celite, and the solvent wasremoved by evaporation. The residue was purified by preparative TLC(MeOH/CHCl₃ 1/9) to give 3 mg (yield: 58%) of Compound 26.

¹ HNMR(DMSO-d₆) δ; 1.173(t,3H,J=7.1 Hz), 1.454(s,3H), 2.300(s,3H),3.064(t,2H,J=7.7 Hz), 3.332(s,3H), 4.077(q,2H,J=7.1 Hz), 5.020(s,1H),6.668(t,1H,J=3.6 Hz), 7.264-7.994(m,6H), 9.144(d,1H,J=1.3 Hz)

Fab-MS(m/z); 581(M+1)⁺

Example 25

Compound 27

To a solution of 60 mg (0.118 mmol of 2-methyl-17-formylstaurosporine ina mixture of 3 ml of THF and 2 ml of MeOH, was added 0.5 ml of anaqueous solution of 107 mg (0.59 mmol) of 2-hydrazino-2-imidazolinhydrobromide, and the mixture was adjusted to pH 2-4 with 3Nhydrochloric acid, and stirred at room temperature for 2 hours. Thereaction mixture was diluted with CHCl₃ and washed succesively with asaturated aqueous solution of NaHCO₃, water and brine, and then driedover K₂ CO₃. The solvent was removed by evaporation and the residue waspurified by preparative TLC (28% NH₄ OH/MeOH/CHCl₃ 0.5/5/95) and treatedwith 0.8N HCl/AcOEt to give 28 mg (36%) of Compound 27.

¹ HNMR(DMSO-d₆) δ; 2.093(m,1H), 2.294(s,3H), 2.671(br.s,3H),3.759(s,3H), 4.035(m,1H), 4.593(br.s,1H), 5.105(s,2H),7.006(dd,1H,J=3.3,9.4 Hz), 7.403-8.243(m,6H), 8.459(s,1H),9.447(d,1H,J=1.7 Hz), 12.551(s,1H).

Fab-MS(m/z); 591(M+1)⁺

Example 26

Compound 28

Compound 28 was obtained from 60 mg (0.118 mol) of2-methyl-17-formylstaurosporine and 57 mg (0.59 mmol) ofdimethylhydrazine hydrochloride by the procedure analogous to that ofExample 25 (yield:30 mg, 46%).

¹ HNMR(DMSO-d₆) δ;1.453(br.s,3H), 2.309(s,3H), 2.928(s,6H),4.072(d,1H,J=3.4 Hz), 5.034(s,2H), 6.703(br.s,1H), 7.269-7.994(m,7H),9.351(d,1H,J=1.7 Hz).

Fab-MS(m/z); 551(M+1)⁺

Example 27

Compound 29

To a solution of 60 mg (0.117 mmol) of Compound 6 and 120 mg (1.17 mmol)of 1H-1,2,4-triazole-3-thiol in 3 ml of CHCl₃ was added 543 mg (2.34mmol) of camphorsulfonic acid, followed by stirring at room temperaturefor 3.5 hours. The reaction mixture was diluted with CHCl₃ and washedwith a saturated aqueous solution of NaHCO₃ and brine, and then driedover K₂ CO₃. The solvent was removed by evaporation and the residue waspurified by silica gel column chromatography (28% NH₄ OH/MeOH/CHCl₃0.5/5/95) and treated with 0.8N HCl/AcOEt to give 41 mg (53%) ofCompound 29.

¹ HNMR(DMSO-d₆) δ;1.458(br.s,3H), 2.294(s,3H), 4.093(br.s,1H),4.581(s,2H), 5.037(s,2H), 6.704(br.s,1H), 7.279-8.020(m,6H),8.429(br.s,1H), 9.307(s,1H), 14. 051(br.s,1H).

Fab-MS(m/z); 594(M+1)⁺

Example 28

Compound 30

Compound 30 was obtained from 60 mg (0.117 mol) of Compound 6 and 165 mg(1.17 mmol) of 2-dimethylaminoethanethiol hydrochloride by the procedureanalogous to that of Example 27 (yield:51 mg, 73%).

¹ HNMR(DMSO-d₆) δ;1.446(s,3H), 2.105(s,6H), 2.305(s,3H), 3.955(s,2H),4.068(d,1H,J=3.4 Hz), 5.027(s,2H), 6.691(t,1H,J=3.5 Hz),7.261-7.993(m,6H), 9.231(d,1H,J=1.3 Hz).

Fab-MS(m/z); 598(M+1)⁺

Example 29

Compound 31

Compound 31 was obtained from 60 mg (0.117 mol) of Compound 6 and 0.12ml (1.17 mmol) of 2-dimethylaminoethanol by the procedure analogous tothat of Example 27 (yield: 16 mg, 23%).

¹ HNMR(DMSO-d₆) δ;1.439(s,3H), 2.213(s,6H), 2.309(s,3H),3.597(t,2H,J=6.0 Hz), 4.072(d,1H,J=3.7 Hz), 4.646(s,2H), 5.031(s,2H),6.705(t,1H,J=3.5 Hz), 7.268-7.995(m,6H), 9.252(d,1H,J=1.3 Hz).

Fab-MS(m/z); 582(M+1)⁺

Example 30

Compound 32

Compound 32 was obtained from 54 mg (0.1 mmol) of Compound 7 and 0.15 ml(2 mmol) of ethanethiol by the procedure analogous to that of Example 27(yield: 46 mg, 73%).

¹ HNMR(DMSO-d₆) δ;1.222(t,3H,J=7.3 Hz), 1.245(t,3H,J=7.3 Hz),2.294(s,3H), 3.271(s,3H), 3.946(s,2H), 3.759(s,2H), 4.061(d,1H,J=3.4Hz), 5.015(s,2H), 6.686(t,1H,J=3.4 Hz), 7.368-7.931(m,5H),9.222(d,1H,J=1.2 Hz).

Fab-MS(m/z); 629(M+1)⁺

Example 31

Compound 33

Compound 33 was obtained from 54 mg (0.1 mmol) of Compound 7 and 283 mg(2 mmol.) of 2-dimethylaminoethanethiol hydrochloride by the procedureanalogous to that of Example 27 (yield: 17 mg, 27% ).

¹ HNMR(DMSO-d₆) δ;1.457(s,3H), 2.116(s,6H), 2.300(s,3H), 3.954(s,2H),4.057(d,1H,J=3.4 Hz), 4.688(s,2H), 5.018(s,2H), 5.158(t,1H,J=5.7 Hz),6.689(t,1H,J=3.5 Hz), 7.366-7.935(m,5H), 9.226(d,1H,J=1.3 Hz).

Fab-MS(m/z); 628(M+1)⁺

Example 32

Compound 34

To a solution of 432 mg (0.87 mmol) of 2-methyl-3-oxostaurosporine in 20ml of dichloroethane was added 0.63 ml (7 mmol) of dichloromethyl methylether and 0.6 ml (4.4 mmol) of titanium tetrachloride and the mixturewas stirred at room temperature for 1.5 hours. The reaction mixture wasdiluted with CHCl₃ and poured to a saturated aqueous solution of NaHCO₃and the organic layer was separated, washed with brine, and then driedover Na₂ SO₄. The solvent was removed by evaporation to give 1.93 g of5,17-diformyl derivative as a crude product.

Compound 34 was obtained from 15 mg of the resulting 5,17-diformylderivative by a procedure analogous to that of Example 1 (yield: 7 mg,46%).

¹ HNMR(DMSO-d₆) δ;1.416(br.s,3H), 2.324(s,3H), 4.079(d,1H,J=3.4 Hz),4.682(d,2H,J=5.6 Hz), 4.721(d,2H,J=5.8 Hz), 5.152(t,1H,J=5.7 Hz),5.222(t,1H,J=5.8 Hz), 6.718(br.s,1H), 7.452-7.969(m,4H), 9.061(s,1H),9.173(s,1H).

Fab-MS(m/z); 555(M+1)⁺

Example 33

Compound 35

11-Cbz derivative of Compound 35 was obtained from 150 mg (0.23 mmol) of2-methyl-11-Cbz-5,17-dihydroxystaurosporine, 162 μl (1.16 mmol) oftriethylamine and 209 μl (1.16 mmol) of pelargonoyl chloride by theprocedure analogous to that of Example 13 (yield: 193 mg, 90%).

¹ HNMR(DMSO-d₆,90°C.) δ; 0.841-0.901(m,6H), 1.256-1.519(m,20H),1.692-1.773(m,4H), 2.322(s,3H), 2.610-2.658(m,4H), 2.670(s,3H),2.749(s,3H), 3.245(s,3H), 4.239(s,1H), 4.649-4.678(m,1H), 5.023(s,2H),5.181(d,1H,J=12.2 Hz), 5.229(d,1H,J=12.5 Hz), 6.974(dd,1H, J=6.0,8.7Hz), 7.201-7.253(m,2H), 7.331-7.456(m,5H), 7.613(d,1H,J=8.8 Hz),7.746(d,1H,J=2.4 Hz), 7.934(d,1H,J=9.0 Hz), 9.006(d,1H,J=2.4 Hz).

Fab-MS(m/z);927(M+1)⁺

Compound 35 was obtained from 170 mg (0.18 mmol) of the 11-Cbzderivative of Compound 35 and 100 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 138 mg, 91%). ¹ HNMR(DMSO-d₆) δ;0.843-0.899(m,6H), 1.244-1.483(m,20H), 1.684-1.749(m,4H),2.071-2.120(m,1H), 2.276(s,3H), 2.648-2.678(m,4H), 3.237(s,3H),4.024(br.s,1H), 4.510(s,1H), 5.020(dd,2H,J=18.1,25.7 Hz),6.916(dd,1H,J=3.0,9.3 Hz), 7.261(dd,1H,J=2.4,9.0 Hz),7.293(dd,1H,J=2.4,9.1 Hz), 7.573(d,1H,J=8.8 Hz), 7.770(d, 1H,J=2.4 Hz),8.093(d,1H,J=9.1 Hz), 8.993(br.s,1H), 9.011(d,1H,J=2.4 Hz),9.237(br.s,1H).

Fab-MS(m/z);793(M+1)⁺

Example 34

Compound 36

11-Cbz derivative of Compound 36 was obtained from 150 mg (0.23 mmol) of2-methyl-11-Cbz-5,17-dihydroxystaurosporine, 432 μl (3.10 mmol) oftriethylamine and 176 mg (3.10 mmol) of 3,6,9,10-tetraoxatridecanoylchloride by the procedure analogous to that of Example 13 (yield: 162mg, 66%).

¹ HNMR(DMSO-d₆,90°C.) δ; 2.319(s,3H), 2.669(s,3H), 2.754(s,3H),3.253(s,3H), 3.390(s,6H), 3.563-3.837(m,24H), 4.133(s,2H), 4.175(s,2H),4.236(s,1H), 4.652-4.698(m,1H), 5.020(s,2H), 5.203(d,1H,J=12.4 Hz),5.243(d,1H,J=12.5 Hz), 6.973(dd,1H,J=6.0,8.6 Hz), 7.209(dd,1H,J=2.4,8.9Hz), 7.258(dd,1H,J=2.2,9.0 Hz), 7.344-7.456(m,5H), 7.610(d,1H,J=9.0 Hz),7.749(d,1H,J=2.2 Hz), 7.933(d,1H,J=9.0 Hz), 9.023(d,1H,J=2.2 Hz).

Fab-MS(m/z);1055(M+1)⁺

Compound 36 was obtained from 150 mg (0.14 mmol) of the 11-Cbzderivative of Compound 36 and 100 mg of 10% Pd/C by the procedureanalogous Co that of Example 8 (yield: 52 mg, 40%).

¹ HNMR(DMSO-d₆) δ; 1.603(br,s,1H), 2.248(s,3H), 3.233(s,3H),3.254(s,3H), 3.420(s,6H), 3.801-3.542(m,24H), 4.107(s,2H), 4.132(s,2H),5.009(s,2H), 6.878(dd,1H,J=3.0,8.9 Hz), 7.161(dd,1H,J=2.4,9.1 Hz),7.193(dd,1H,J=2.3,9.0 Hz), 7.472(d,1H,J=2.4 Hz), 7.564(d,1H,J=8.9 Hz),8.097(d,1H,J=9.0 Hz), 9.008(d,1H,J=2.2 Hz).

Fab-MS(m/z);921(M+1)⁺

Example 35

Compound 37

11-Cbz derivative of Compound 37 was obtained from 150 mg (0.23 mmol) ofthe 11-Cbz derivative of Compound 16, 81 μl (0.58 mmol) of triethylamineand 160 mg (0.58 mmol) of palmitoyl chloride by the procedure analogousto that of Example 13 (yield: 147 mg, 56%).

¹ HNMR(DMSO-d₆, 90°C.) δ;0.823-0.874(m,6H), 1.124-1.411(m,48H),1.621-1.695(m,4H), 2.281(s,3H), 2.329-2.382(m,4H), 2.627(s,3H),2.740(s,3H), 3.282(s,3H), 4.167(s,1H), 4.635-4.686(m,1H), 4.974(s,2H),5.175(d,1H,J=12.5 Hz), 5.227(d,1H,J=12.5 Hz), 6.895(dd,1H,J=5.9,8.5 Hz),7.342-7.447(m,5H), 7.472(d,1H,J=8.8 Hz), 7.660(dd,1H,J=2.0,9.0 Hz),7.791-7.822(m,2H), 8.336(d,1H,J=2.0 Hz), 9.240(d,1H,J=2.0 Hz),9.631(s,1H), 9.692(s,1H).

Fab-MS(m/z);1121(M+1)⁺

Compound 37 was obtained from 147 mg (0.13 mmol) of the 11-Cbzderivative of Compound 37 and 75 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 114 mg, 85%).

¹ HNMR(DMSO-d₆) δ; 0.864-0.898(m,6H), 1.267-1.371(m,48H),1.665-1.719(m,4H), 2.071-2.142(m,1H), 2.328(s,3H), 2.379-2.432(m,4H),2.509(s,3H), 2.731(s,3H), 4.072(br.s,1H), 4.451(s,1H), 5.053(s,2H),6.935(dd,1H,J=3.1,9.4 Hz), 7.516(d,1H,J=8.8 Hz), 7.743(dd,1H,J=2.1,9.2Hz), 7.976(dd,1H,J=2.0,8.8 Hz), 8.032(d,1H,J=9.3 Hz), 8.485(d,1H,J=2.0Hz), 8.915(br.s,2H), 9.311(d,1H,J=2.2 Hz), 10.018(s,1H), 10.085(s,1H).

Fab-MS(m/z);987(M+1)⁺

Example 36

Compound 38

11-Cbz derivative of Compound 38 was obtained from 133 mg (0.21 mmol) ofthe 11-Cbz derivative of Compound 16, 72 μl (0.51 mmol) of triethylamineand 93 μl (0.51 mmol) of pelargonoyl chloride by the procedure analogousto that of Example 13 (yield: 79 mg, 42%).

¹ HNMR(DMSO-d₆, 90° C.) δ; 0.843-0.889(m,6H), 1.255-1.419(m,20H),1.624-1.717(m,4H), 2.283(s,3H), 2.332-2.385(m,4H), 2.628(s,3H),2.741(s,3H), 3.283(s,3H), 4.169(s,1H), 4.635-4.686(m,1H), 4.978(s,2H),5.176(d,1H,J=12.5 Hz), 5.228(d,1H,J=12.5 Hz), 6.899(dd,1H,J=5.7,8.7 Hz),7.326-7.449(m,6H), 7.477(d,1H,J=8.8 Hz), 7.661(dd,1H,J=2.2,9.0 Hz),7.807(d,1H,J=9.3 Hz), 8.336(d,1H,J=2.0 Hz), 9.242(d,1H,J=2.0 Hz),9.635(s,1H), 9.697(s,1H).

Fab-MS(m/z);925(M+1)⁺

Compound 38 was obtained from 79 mg (0.086 mmol) of the 11-Cbzderivative of Compound 38 and 40 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 60 mg, 85%).

¹ HNMR(DMSO-d₆) δ; 0.855-0.886(m,6H), 1.237-1.331(m,20H),1.607-1.679(m,4H), 2.028-2.097(m,1H), 2.296(s,3H), 2.339-2.393(m,4H),2.466(s,3H), 2.674(s,3H), 4.008(br.s,1H), 4.456(s,2H), 5.014(s,2H),6.895(dd,1H,J=2.9,9.3 Hz), 7.474(d,1H,J=9.0 Hz), 7.697(dd,1H,J=2.0,9.3Hz), 7.926(dd,1H,J=2.0,8.8 Hz), 7.990(d,1H, J=9.0 Hz), 8.455(d,1H,J=2.0Hz), 8.943(br.s,1H), 9.179(br.s,1H), 9.274(d,1H,J=2.0 Hz), 9.979(s,1H),10.064(s,1H).

Fab-MS(m/z);791(M+1)⁺

Example 37

Compound 39

A solution of 200 mg (0.31 mmol) of2-methyl-11-Cbz-5,17-dihydroxystaurosporine in 6 ml of DMF was cooled to0° C. and 37 mg (0.93 mmol) of 60% sodium hydride was added thereto,followed by stirring for 30 minutes. To the reaction mixture was added473 μl (1.55 mmol) of 1-bromohexadecane, followed by stirring for 3hours. After the reaction was stopped by an addition of a saturatedammonium chloride aqueous solution, the reaction mixture was extractedwith AcOEt. The extract was washed with brine, and dried over Na₂ SO₄.The solvent was removed by evaporation, and the residue was purified bysilica gel column chromatography (MeOH/CHCl₃ 1/200) to give 228 mg (82%)of 11-Cbz derivative of Compound 39.

¹ HNMR(CDCl₃) δ; 0.876(t,6H,J=6.8 Hz), 1.262-1.541(m,52H),1.837-1.894(m,4H), 2.150(br.s,1H), 2.425(s,3H), 2.509(s,3H),2.540-2.592(m,1H), 2.799(s,3H), 3.395(s,3H), 3.999(br.s,1H),4.103(t,2H,J=6.6 Hz), 4.224(t,2H,J=6.4 Hz), 4.835(br.s,1H),5.170(d,2H,J=12.0 Hz), 6.685(br.s,1H), 7.061(dd,1H,J=2.4,9.0 Hz),7.108-7.161(m,2H), 7.287-7.595(m,7H), 9.080(s,1H).

Fab-MS(m/z);1095(M+1)⁺

Compound 39 was obtained from 211 mg (0.23 mmol) of the 11-Cbzderivative of Compound 39 and 70 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 126 mg, 67%).

¹ HNMR(CDCl₃) δ; 0.879(t,6H,J=6.8 Hz), 1.204-1.570(m,52H),1.612(br.s,3H), 1.817-1.907(m,4H), 2.308(s,3H), 2.363-2.409(m,1H),2.685(dd,1H,J=3.9,14.9 Hz), 3.347(br.s,3H), 3.391(s,3H), 3.857(br.s,1H),4.106(t,2H,J=6.6 Hz), 4.227(t,2H,J=6.3 Hz), 4.915(d,2H,J=3.9 Hz),6.495(d,1H,J=4.9 Hz), 7.035(dd,1H,J=2.7,9.3 Hz), 7.106(dd,1H,J=2.4,8.5Hz), 7.154(d,1H,J=8.8 Hz), 7.327(d,1H,J=2.7 Hz), 7.776(d,1H,J=9.3 Hz),9.070(d,1H,J=2.4 Hz).

Fab-MS(m/z);961(M+1)⁺

Example 38

Compound 40

11-Cbz derivative of Compound 40 was obtained from 200 mg (0.31 mmol) of2-methyl-11-Cbz-5,17-dihydroxystaurosporine, 37 mg (0.93 mmol) of 60%sodium hydride, and 296 μl (1.55 mmol) of 1-bromononane by the procedureanalogous to that of Example 37 (yield: 278 mg, 72%).

¹ HNMR(DMSO-d₆,70° C.) δ; 0.843-0.882(m,6H), 1.278-1.497(m,24H),1.765-1.834(m,4H), 2.254(s,3H), 2.624(s,3H), 2.732(s,3H), 3.207(s,3H),4.086(t,2H,J=6.5 Hz), 4.134(t,2H,J=6.4 Hz), 4.161(br.s,1H),4.654(br.s,1H), 5.040(d,2H,J=2.2 Hz), 5.170(d,1H,J=12.5 Hz),5.226(d,1H,J=12.2 Hz), 6.881(dd,1H,J=5.9,8.8 Hz), 7.075-7.108(m,2H),7.353-7.460(m,7H), 7.790(d,1H,J=8.8 Hz), 8.930(d,1H,J=2.7 Hz)

Fab-MS(m/z);899(M+1)⁺

Compound 40 was obtained from 250 mg (0.23 mmol) of the resulting 11-Cbzderivative of Compound 40 and 100 mg of 10% Pd/C by the procedureanalogous to that of Example 8 (yield: 182 mg, 80%).

¹ HNMR(DMSO-d₆) δ; 0.850-0.884(m,6H), 1.224-1.514(m,24H),1.769-1.838(m,4H), 2.005-2.079(m,1H), 2.272(s,3H), 2.447(s,3H),2.664(s,3H), 3.265(s,3H), 3.990(br.s,3H), 4.073(t,2H,J=6.6 Hz),4.131(t,2H,J=6.7 Hz), 4.451(s,1H), 5.069(s,2H), 6.863(dd,1H,J=3.3,9.4Hz), 7.114-7.155(m,2H), 7.446(d,1H,J=8.8 Hz), 7.468(d,1H,J=2.7 Hz),7.947(d,1H,J=9.5 Hz), 8.940(d,1H,J=2.7 Hz), 8.965(br.s,1H),9.170(br.s,1H)

Fab-MS(m/z); 765(M+1)⁺

Example 39

Injections

Compound 22 (2.0 g) was dissolved in 20 l of ethanol, and the solutionwas subjected to pressure filtration through Millipore Filter (poresize: 0.22μ) for sterilization. The resulting sterile filtrate was putinto brown vials in 5.0 ml portions, and lyophilized in a conventionalmanner to give 0.5 mg/vial of lyophilized preparations.

Example 40

Tablets

Tablets were prepared in a conventional manner using 180 mg of Compound22, 90 mg of lactose, 40 mg of cornstarch, 4 mg of polyvinyl alcohol, 28mg of Avicel and 1 mg of magnesium stearate.

Although the invention has been described in specific terms, it is to beunderstood that the described embodiments are only illustrative andvarious change and modifications may be imparted thereto withoutdeparting from the scope of the invention which is limited solely by theappended claims.

We claim:
 1. An indolocarbazole derivative represented by the formula(I): ##STR24## {wherein R¹ represents substituted or unsubstituted loweralkyl, R² represents hydrogen, substituted or unsubstituted lower alkyl,R³ and R⁴ are the same or different, and represent hydrogen, nitro, NR⁵R⁶ (wherein R⁵ and R.sup. 6 are the same or different and representhydrogen, lower alkyl, an amino acid residue in which the hydroxy groupis removed from the carboxylic acid, alkanoyl or CO(CH₂)_(m) NR⁷ R⁸wherein m is an integer of 0 to 3, R⁷ and R⁸ are the same or differentand represent hydrogen or lower alkyl, or R⁷ and R⁸ are combinedtogether with nitrogen atom to form a heterocyclic ring, OR⁹ (wherein R⁹represents long-chain alkyl, alkanoyl, COCH₂ (OCH₂ CH₂)_(k) OCH₃ whereink is an integer of 1 to 6, or CONR⁷ R⁸, substituted or unsubstitutedlower alkyl, substituted or unsubstituted lower alkenyl, or CH═NNR¹⁰ R¹¹wherein R¹⁰ and R¹¹ are the same or different and represent hydrogen,lower alkyl or a member selected from the group consisting of pyrrolyl,imidazolyl, thienyl, furyl, triazolyl, tetrazolyl, thiazolyl, oxazolyl,pyridyl, pyrimidinyl, indolyl and quinolyl, W¹ and W² are both hydrogenor are combined together to represent oxygen, and Me represents methyl,provided that when R¹ is unsubstituted lower alkyl, R³ and R⁴ are notsimultaneously hydrogen}, or a pharmaceutically acceptable salt thereof.2. A compound according to claim 1, in which R¹ is unsubstituted loweralkyl.
 3. A compound according to claim 2, in which R¹ is methyl.
 4. Acompound according to any of claims 1-3, in which R² is hydrogen.
 5. Acompound according to any of claims 1-3, in which R³ is NR⁵ R⁶ (whereinR⁵ and R⁶ are the same or different and represent hydrogen, lower alkyl,an amino acid residue where hydroxy group in the carboxylic acid isremoved from the amino acid, alkanoyl or CO(CH₂)_(m) NR⁷ R⁸ (wherein mis an integer of 0 to 3, R⁷ and R⁸ are the same or different andrepresent hydrogen or lower alkyl, or R⁷ and R⁸ are combined togetherwith nitrogen atom to form a heterocyclic ring)), OR⁹ (wherein R⁹represents long chain alkyl, alkanoyl, COCH₂ (OCH₂ CH₂)_(k) OCH₃(wherein k is an integer of 1 to 6), or CONR⁷ R⁸ (wherein R⁷ and R⁸ havethe same meanings as defined above)), substituted or unsubstituted loweralkyl, substituted or unsubstituted lower alkenyl, or CH═NNR¹⁰ R¹¹(wherein R¹⁰ and R¹¹ are the same or different and represent hydrogen,lower alkyl or a heterocyclic ring containing nitrogen atom).
 6. Acompound according to any of claims 1-3, in which W represents twohydrogen atoms.
 7. A pharmaceutical composition which comprises, as anactive ingredient, a compound as defined by claim 1, and apharmaceutically acceptable carrier.
 8. A method of treatingthrombocytopenia by administering, to a patient suffering fromthrombocytopenia, an effective amount of the compound according toclaim
 1. 9. A compound according to claim 4, in which R³ is NR⁵ R⁶. 10.A compound according to claim 4, in which W represents two hydrogenatoms.
 11. A compound according to claim 9, in which W represents twohydrogen atoms.